Prestige Antibodies® in Breast Cancer Research

The Human Protein Atlas



proteinatlas.org

The Human Protein Atlas is Characterizing the Human Proteome

The Human Protein Atlas (HPA) project was initiated in 2003 by Swedish researchers, headed by Professor Mathias Uhlén, and funded by the Knut and Alice Wallenberg Foundation.1,2 It is a unique world leading effort performing systematic exploration of the human proteome using antibodies.

The aim of the HPA project is to present an expression map of the complete human proteome. To accomplish this, highly specific Triple A polyclonal antibodies are developed to all protein coding human genes and protein profiling is established in a multitude of tissues and cells using tissue arrays. Applications applied are immunohistochemistry (IHC), Western blot (WB) analysis, protein array assay and immunofluorescent based confocal microscopy (ICC-IF).

The Human Protein Atlas, November 2014

The 13th version of the Human Protein Atlas, released in November 2014, presents a tissue-based map of the complete human proteome. The extensive amount of data is divided into four separate 'sub atlases': the Tissue Atlas, the Cancer Atlas, the Subcell Atlas and the Cell Line Atlas. For all proteins represented in the Tissue Atlas, the expression profiles are based on IHC analysis on a large number of human tissues. The presentation of protein expression data in correlation to RNA sequencing data for each gene has now been included. In the Cancer Atlas, differentially expressed genes in several cancers can be studied, while the Subcell Atlas presents subcellular localization by confocal microscopy. Additional information about protein expression in common cell lines is included in the Cell Line Atlas, which has become an appreciated toolbox for research.

Tissue microarrays containing samples from 48 different normal human tissues, 20 different cancer types and 44 different human cell lines are utilized within the project. The 48 normal tissues are present in triplicate samples and represent 82 different cell types. All normal tissue images have undergone pathology-based annotation of expression levels and are displayed on the normal Tissue Atlas presenting information regarding the expression profiles of human genes both on mRNA and protein level. The mRNA expression data is derived from deep sequencing of RNA (RNA-Seq) from 27 major different normal tissue types.

The Cancer Atlas contains gene expression data based on protein expression patterns in a multitude of human cancer specimens. Altogether 216 different cancer samples, corresponding to the 20 most common forms of human cancer, have been analyzed for all included genes. All cancer tissue images have been manually annotated by pathologists and just as for the normal Tissue Atlas, protein data includes protein expression levels corresponding to 16.621 genes for which there are available antibodies.

Validation in Breast Tissue Samples and Cell Lines

IHC images from normal breast samples from three different individuals are available for each antibody in the normal Tissue Atlas. In addition, for each antibody, breast tumor samples from up to 12 patients in duplicates are presented in the Cancer Atlas and for the majority of the antibodies, also images from the MCF-7 and SK-BR-3 breast cell lines in the Cell Line Atlas.

References

  1. Uhlén M et al. (2010) Towards a knowledge-based Human Protein Atlas. Nat Biotechnol 28(12):1248–50.
  2. Uhlén et al. (2015) Proteomics. Tissue-based map of the human proteome. Science 23;347(6220).

Prestige Antibodies® Powered by Atlas Antibodies

Prestige Antibodies – the Building Blocks of HPA

The uniqueness and low cross reactivity of Prestige Polyclonals to other proteins are due to a thorough selection of antigen regions, affinity purification on the recombinant antigen, validation using several methods and a stringent approval process.

Development

The Prestige Antibodies are developed against recombinant human Protein Epitope Signature Tags (PrESTs) of approximately 50 to 150 amino acids. These protein fragments are designed, using a proprietary software, to contain unique epitopes present in the native protein suitable for triggering the generation of antibodies of high specificity. This is achieved by a complete human genome scanning to ensure that PrESTs with the lowest homology to other human proteins are used as antigens.

Approval

The approval of the Prestige Antibodies relies on a combined validation of the experimental results using IHC, WB or ICC-IF, from RNA sequencing and from information obtained via bioinformatics prediction methods and literature. Since the literature is often inconclusive, an important objective of the HPA project has been to generate paired antibodies with non-overlapping epitopes towards the same protein target, allowing the results and validation of one antibody to be used to validate the other one.

Prestige Antibodies Catalog

Today, there are more than 17,000 Prestige Polyclonals with 2,000 new antibodies added each year.

The antibodies developed and characterized within the Human Protein Atlas project are made available to the scientific community by Sigma-Aldrich as Prestige Antibodies and Atlas Antibodies under the brand name Triple A Polyclonals.

Monoclonal Antibody Development

Prestige Antibodies also include a selected number of mouse monoclonal antibodies. The monoclonal catalog is regularly expanding with new products every year.

Unique Features

Special care is taken in offering clones recognizing only unique nonoverlapping epitopes and/or isotypes. Using the same stringent PrEST production process and characterization procedure as for the Triple A Polyclonals, the monoclonal antibodies offer outstanding performance in approved applications, together with defined specificity, secured continuity and stable supply. In general they also permit high working dilutions and contribute to more standardized assay procedures.

Clone Selection

Functional characterization is performed on a large number of ELISA positive cell supernatants to select the optimal clones for each application prior to subcloning and expansion of selected hybridomas.

Epitope Mapping

Clones are epitope-mapped using synthetic overlapping peptides in a bead-based array format for selection of clones with nonoverlapping epitopes only.

Isotyping

All monoclonal antibodies are isotyped to allow for multiplexing using isotype-specific secondary antibodies.

Hybridoma Cell Cultivation

Atlas Antibodies use in vitro methods for the production scale-up phase thus replacing the use of mice for production of ascites fluid.

Antibody Characterization

The characterization of Prestige Monoclonal Antibodies starts with an extensive literature search to select the most relevant and clinically significant tissues to use for IHC characterization. Often there are more than one tissue type displayed in the IHC application data for each antibody. In addition to positive stained tissue, a negative control tissue staining is also displayed and if relevant, clinical cancer tissue staining.

The Western blot (WB) characterization includes results from endogenous human cell or tissue protein lysates or optionally recombinant full-length human protein lysates.

Each monoclonal antibody is thus supplied with the most relevant characterization data for its specific target.

The product numbers of all Prestige Polyclonals start with ”HPA” and of monoclonal antibodies with “AMAB”.

Clinical Markers (ESR1, HER2, Ki67, PGR)

Established Clinical Breast Cancer Markers

 

Cat. No.
Target Protein Product Description Validated Applications
HPA000449
Estrogen receptor Anti-ESR1 IHC,WB
HPA000450 Estrogen receptor Anti-ESR1 IHC,WB
HPA004751 Progesteron receptor Anti-PGR1 IHC,ICC-IF
HPA008428 Progesteron receptor Anti-PGR IHC
HPA017176 Progesteron receptor Anti-PGR IHC
HPA001383 HER2/ERBB2 Anti-ERBB2 IHC,WB
AMAb90627 HER2/ERBB2 Anti-HER2 IHC,WB
HPA000451 Ki67/MKI67 Anti-MKI672 IHC,ICC-IF
HPA001164 Ki67/MKI67 Anti-MKI673 IHC,ICC-IF
AMAb90870 Ki67/MKI67 Anti-MKI67 IHC
  1. Pereira CB et al. Prognostic and Predictive Significance of MYC and KRAS Alterations in Breast Cancer from Women Treated with Neoadjuvant Chemotherapy. PLoS One 2013;8(3):e60576.
  2. Camilleri M et al. Neuropeptide S receptor induces neuropeptide expression and associates with intermediate phenotypes of functional gastrointestinal disorders. Gastroenterology 2010 Jan;138(1):98–107.e4.
  3. Roca H et al. IL-4 induces proliferation in prostate cancer PC3 cells under nutrient-depletion stress through the activation of the JNK-pathway and survivin upregulation. J Cell Biochem 2012 May; 113(5):1569–1580.


HER2/ERBB2

Immunohistochemical staining of human breast tumour using Anti-HER2 (AMAb90627) shows strong membranous (combined with moderate cytoplasmic) positivity in tumour cells in HER2-positive ductal carcinoma, while HER2-negative ductal carcinoma shows no membranous positivity. By Western Blot analysis, HER2 is detected in the breast cancer cell line SK-BR-3.

Progesteron Receptor

IHC staining using the Anti-PGR antibody (HPA004751) in normal human corpus (uterine) tissue shows strong nuclear positivity in glandular cells. In the presented breast cancer sample, the staining of tumor cells is also nuclear. ICC-IF shows nuclear staining in U-251MG cells.

Estrogen Receptor

The Anti-ESR1 antibody (HPA000449) shows distinct nuclear positivity in glandular cells in human breast tissue and in tumor cells in breast cancer samples using IHC.

 

IHC staining using the Anti-ESR1 antibody (HPA000450) shows strong nuclear positivity in glandular and stromal cells of human corpus, uterine tissue and in tumor cells in breast cancer.

Ki67

The Anti-MKI67 antibody (HPA000451) shows strong nuclear positivity in a fraction of cells in the reaction center in human lymph node using IHC. In breast cancer, the staining of tumor cells is also nuclear and by ICC-IF, staining of the human cell line U-2OS shows positivity in nucleoli.

 

IHC staining of human tonsil tissue using the Anti-MKI67 antibody (HPA001164) shows nuclear staining of reaction center cells. In tumor cells in breast cancer, the staining is mainly nuclear and in U-2OS cells, using ICC-IF, nucleoli show strong positivity.

 

IHC staining of lymph node in human colon shows strong nuclear and nucleolar immunoreactivity in the reaction centrum cells using the monoclonal Anti-MKI67 antibody (AMAb90870). In uterus, nuclear positivity in a subset of glandular cells is shown.

Antibodies used in Breast Cancer Research

In this section, antibodies are selected either on a reference/article-basis or on breast cancer relevance for the corresponding target protein.
 

BRCA1

The Anti-BRCA1 antibody (HPA034966) shows positivity in glandular cells in normal human breast tissue and in tumor cells in breast cancer samples using IHC.

IHC staining using the Anti-BRCA2 antibody (HPA026815) in normal human breast tissue shows positivity in glandular cells. In breast cancer, nuclear staining of tumor cells is shown.

ACAT1

Immunohistochemical staining of human liver tissue using Anti-ACAT1 (HPA004428) shows strong cytoplasmic positivity in hepatocytes. By Western Blot analysis, ACAT1 is detected in the human cell lines RT-4 and U251-MG and in liver and tonsil tissue lysates. By ICC-IF in the human cell line A-431, positivity is shown in mitochondria.

CD44

Immunohistochemical staining of human esophagus tissue using Anti-CD44 (HPA005785) shows strong strong cytoplasmic and membranous positivity in squamous epithelial cells. By Western Blot analysis, CD44 is detected in the human cell line U-251MG. ICC-IF in the human cell line U-251MG shows positivity in plasma membrane.

 

Cat. No. Target Protein Product Description Validated Applications
HPA008788 53BP1 Anti-TP53BP1 IHC,ICC-IF
HPA022133 53BP1 Anti-TP53BP1 IHC,WB*,ICC-IF
HPA004428 ACAT1 Anti-ACAT11,2 IHC,WB*,ICC-IF
HPA007569 ACAT1 Anti-ACAT12-4 IHC,WB,ICC-IF
HPA007912 AGR2 Anti-AGR25 IHC,WB
HPA024210 AIB1/NCOA3 Anti-NCOA3 IHC,ICC-IF
AMAb90660 Anillin/ANLN Anti-ANLN IHC,WB
AMAb90662 Anillin/ANLN Anti-ANLN IHC,WB
HPA005680 Anillin/ANLN Anti-ANLN6 IHC,WB,ICC-IF
HPA024006 ARG1 Anti-ARG17 IHC,WB
HPA005468 ASAH1 Anti-ASAH18,9 IHC,WB
HPA028264 ATR Anti-ATR IHC
HPA029455 BAAT1/BRAT1 Anti-BRAT1 IHC,WB
HPA003175 BACH1 Anti-BACH110 IHC,WB,ICC-IF
HPA028814 BAP1 Anti-BAP1 IHC,WB,ICC-IF
HPA044864 BARD1 Anti-BARD1 IHC,ICC-IF
HPA029159 Beta-Catenin Anti-CTNNB1 IHC,WB*,ICC-IF
HPA029160 Beta-Catenin Anti-CTNNB1 IHC, IF
HPA002317 BIRC3/API2 Anti-BIRC311 IHC,WB,ICC-IF
HPA012897 BIT1/ PTRH2 Anti-PTRH212,13 IHC,WB,ICC-IF
HPA005689 Blooms Syndrome Prot Anti-BLM IHC,ICC-IF
HPA030472 Bmi1 Anti-BMI1 IHC,WB*,ICC-IF
HPA034966 BRCA1 Anti-BRCA1 IHC
HPA026815 BRCA2 Anti-BRCA2 IHC,ICC-IF
HPA005474 BRIP1/FANCJ Anti-BRIP1 IHC,WB,ICC-IF
HPA036596 C11orf51/ANAPC15 Anti-ANAPC15 IHC,WB,ICC-IF
HPA051365 CAR/NR1I3 Anti-NR1I3 IHC,ICC-IF
HPA001302 CASP8 Anti-CASP8 IHC,WB,ICC-IF
HPA005688 CASP8 Anti-CASP8 IHC,WB,ICC-IF
HPA008773 CAXII/CA12 Anti-CA1214-17 IHC,WB
HPA005785 CD44 Anti-CD4418-22 IHC,WB,ICC-IF
HPA028900 CD82 Anti-CD82 IHC,WB
HPA004812 CDH1 Anti-CDH1 IHC,ICC-IF
HPA019758 CEA/CEACAM5 Anti-CEACAM5 IHC,WB
HPA001878 CHEK2 Anti-CHEK2 IHC,WB
HPA001254 CKB Anti-CKB IHC
HPA004135 CRABP2 Anti-CRABP2 IHC,WB,ICC-IF
HPA015955 CTNND1 Anti-CTNND1 IHC,WB*,ICC-IF
HPA010663 CX32/GJB1 Anti-GJB123 IHC,WB
HPA018169 Cyclin E1 Anti-CCNE1 IHC,WB,ICC-IF
HPA020626 Cyklin A2 Anti-CCNA2 IHC,WB
HPA023040 Cytokeratin 14/CK14 Anti-KRT14 IHC,WB*,ICC-IF
HPA000452 Cytokeratin 17/CK17 Anti-KRT1724 IHC,WB
HPA000453 Cytokeratin 17/CK17 Anti-KRT17 IHC,WB,ICC-IF
HPA000258 DACH2 Anti-DACH225 IHC,ICC-IF
HPA019907 DBC1/KIAA1967 Anti-KIAA1967 IHC,WB*,ICC-IF
HPA019943 DBC1/KIAA1967 Anti-KIAA1967 IHC,ICC-IF
HPA022962 DCAF7 Anti-DCAF726 IHC, WB
HPA003315 Decorin/DCN Anti-DCN27,28 IHC, WB
HPA028483 DIRAS3 Anti-DIRAS3 IHC,WB
HPA028557 DIRAS3 Anti-DIRAS3 IHC,WB
HPA029384 DIRAS3 Anti-DIRAS3 IHC
AMAb90816 EGFR Anti-EGFR IHC,WB
AMAb90819 EGFR Anti-EGFR WB
HPA001200 EGFR Anti-EGFR29 IHC
HPA018530 EGFR Anti-EGFR30 IHC,WB,ICC-IF
HPA003901 Endoplasmin/ HSP90B1 Anti-HSP90B127,31 IHC,WB,ICC-IF
HPA002025 ERLIN2 Anti-ERLIN232,33 IHC,WB*,ICC-IF
HPA026676 ERFF/C1orf64 Anti-C1orf6434 IHC,WB,ICC-IF
HPA007425 FAAH Anti-FAAH35 IHC,ICC-IF
HPA035305 FGFR2 Anti-FGRF2 IHC,WB,ICC-IF
HPA029731 GATA3 Anti-GATA3 IHC, WB
HPA009177 GCDFP/PIP Anti-PIP IHC,WB
HPA017046 GEF-H1 Anti-ARHGEF236,37 IHC,WB
HPA025226 GGH Anti-GGH35 IHC,WB
HPA008763 Granulin Anti-GRN38 IHC,ICC-IF, WB
HPA028747 Granulin Anti-GRN38 IHC,ICC-IF
HPA001275 HIF-1 alpha/HIF1A Anti-HIF1A39-42 IHC,ICC-IF
HPA008436 HJURP Anti-HJURP43-45 IHC,WB,ICC-IF
HPA004727 HMGCL Anti-HMGCL2 IHC,WB
HPA008338 HMGCR Anti-HMGCR46 IHC
HPA021467 HSD17B14 Anti-HSD17B14 IHC,WB,ICC-IF
HPA000764 KLK3/PSA Anti-KLK347,48 IHC
HPA019693 LSP1 Anti-LSP1 IHC,WB,ICC-IF
HPA001939 MMP2 Anti-MMP2 IHC
HPA004179 MUC1/CA15-3 Anti-MUC1 IHC,WB
HPA007235 MUC1/CA15-3 Anti-MUC1 IHC
HPA008855 MUC1/CA15-3 Anti-MUC1 IHC,ICC-IF
HPA001429 NBN Anti-NBN IHC,WB,ICC-IF
HPA030278 NRP1 Anti-NRP1 IHC
HPA029814 Oncostatin M Anti-OSM49 IHC,WB
AMAb90956 P53 Anti-P53 IHC,WB
HPA021241 PHGDH Anti-PHGDH50-52 IHC,WB*,ICC-IF
HPA031314 PGD Anti-PGD IHC,WB*
HPA006563 PKC alpha/PKCA Anti-PKCA IHC,WB*,ICC-IF
HPA006564 PKC alpha/PKCA Anti-PKCA IHC,WB*,ICC-IF
HPA003412 PLAT Anti-PLAT IHC,WB,ICC-IF
HPA006366 POLRMT Anti-POLRMT53 IHC
HPA020376 PSPH Anti-PSPH50 IHC,WB
HPA047183 PTMA Anti-PTMA IHC,ICC-IF
HPA008890 PTTG1 Anti-PTTG1 IHC,ICC-IF
HPA037503 RAP80/UIMC1 Anti-UIMC1 IHC,WB
HPA037504 RAP80/UIMC1 Anti-UIMC1 IHC,WB,ICC-IF
HPA006079 REST Anti-REST54,55 IHC,ICC-IF
HPA003624 RBM3 Anti-RBM356,57 IHC,WB*,ICC-IF
AMAb90655 RBM3 Anti-RBM358-65 IHC,WB
HPA009026 RRBP1 Anti-RRBP166 IHC,ICC-IF, WB
HPA035857 rs4973768/SLC4A7 Anti-SLC4A7 IHC
HPA001893 SIX1 Anti-SIX167-74 IHC,WB,ICC-IF
AMAb90544 SIX1 Anti-SIX1 IHC,WB
HPA014404 SNCG Anti-SNCG IHC,WB
HPA017254 STK11 Anti-STK11 IHC,WB,ICC-IF
HPA002830 SURVIvin/BIRC5 Anti-BIRC5 IHC,WB,ICC-IF
HPA000500 T-STAR/KHDRBS3 Anti-KHDRBS375,76 IHC,WB
HPA004823 Tenascin C/TNC Anti-TNC77-79 IHC,WB
HPA003425 TFF1 Anti-TFF180-82 IHC,WB
HPA002982 THBD Anti-THBD IHC,WB
HPA019881 THEM2/ACOT13 Anti-ACOT13 IHC,WB*,ICC-IF
HPA006458 TOP2A Anti-TOP2A IHC,WB,ICC-IF
HPA026773 TOP2A Anti-TOP2A IHC,ICC-IF
HPA014405 UGT8 Anti-UGT883 IHC,ICC-IF
HPA007547 ULBP1 Anti-ULBP184 IHC
HPA023930 ZNF703 Anti-ZNF70332 IHC,ICC-IF

* WB both in human and rodent samples

  1. Sanchez-Alvarez R et al. Ethanol exposure induces the cancer-associated fibroblast phenotype and lethal tumor metabolism: Implications for breast cancer prevention. Cell Cycle 2013 Jan 15; 12(2):289–301.
  2. Martinez-Outschoorn UE et al. Ketone bodies and two-compartment tumor metabolism: Stromal ketone production fuels mitochondrial biogenesis in epithelial cancer cells. Cell Cycle 2012 Nov 1; 11(21):3956–3963.
  3. Martinez-Outschoorn UE et al. Ketone body utilization drives tumor growth and metastasis. Cell Cycle 2012 Nov 1;11(21):3964–71.
  4. Chang HT et al. Ketolytic and glycolytic enzymatic expression profiles in malignant gliomas: implication for ketogenic diet therapy. Nutr Metab (Lond) 1047. Epub 2013/07/05.
  5. Hrstka R et al. AGR2 Predicts Tamoxifen Resistance in Postmenopausal Breast Cancer Patients. Dis Markers 2013; 35(4):207–212. Epub 2013/09/03.
  6. O´Leary PC et al. Systematic antibody generation and validation via tissue microarray technology leading to identification of a novel protein prognostic panel in breast cancer. BMC Cancer. 2013 Apr 2;13:175.
  7. de Boniface J et al. Expression patterns of the immunomodulatory enzyme arginase 1 in blood, lymph nodes and tumor tissue of early-stage breast cancer patients. Oncoimmunology 2012 Nov 1; 1(8):1305–1312.
  8. Lucki NC, Sewer MB. Genistein Stimulates MCF-7 Breast Cancer Cell Growth by Inducing Acid Ceramidase (ASAH1) Gene Expression. J Biol Chem 2011 Jun 3; 286(22):19399–19409. Epub 2011 Apr 14.
  9. Lucki NC et al. Acid Ceramidase (ASAH1) Represses Steroidogenic Factor 1-Dependent Gene Transcription in H295R Human Adrenocortical Cells by Binding to the Receptor. Mol Cell Biol 2012 Nov; 32(21):4419–4431.
  10. Liang Y et al. Transcriptional Network Analysis Identifies BACH1 as a Master Regulator of Breast Cancer Bone Metastasis. J Biol Chem 2012 Sep 28;287(40):33533–44.
  11. Almubarak H et al. Zoledronic acid directly suppresses cell proliferation and induces apoptosis in highly tumorigenic prostate and breast cancers. J Carcinog 2011 Jan 15;10:2.
  12. Brunquell C et al. TLE1 is an anoikis regulator and is downregulated by Bit1 in breast cancer cells. Mol Cancer Res 2012 Nov; 10(11):1482–1495. Epub 2012/09/04.
  13.  Karmali PP et al. Metastasis of tumor cells is enhanced by downregulation of bit1. PLoS One 2011;6(8):e23840.
  14. Vermeulen JF et al. Immunophenotyping invasive breast cancer: paving the road for molecular imaging. BMC Cancer 12240. Epub 2012 Jun 13.
  15. Davidson B et al. Gene expression signatures differentiate ovarian/peritoneal serous carcinoma from breast carcinoma in effusions. J Cell Mol Med 2011 Mar;15(3):535–44.
  16. Vermeulen et al. Differential expression of growth factor receptors and membrane-bound tumor markers for imaging in male and female breast cancer. PLoS One 2013;8(1):e53353.
  17. Tafreshi NK et al. Noninvasive detection of breast cancer lymph node metastasis using carbonic anhydrases IX and XII targeted imaging probes. Clin Cancer Res 2012 Jan 1;18(1):207–19.
  18. Vazquez-Martin A et al. Metformin regulates breast cancer stem cell ontogeny by transcriptional regulation of the epithelial-mesenchymal transition (EMT) status. Cell Cycle 2010 Sep 15;9(18):3807–14.
  19. Baccelli I et al. Identification of a population of blood circulating tumor cells from breast cancer patients that initiates metastasis in a xenograft assay. Nat Biotechnol 2013 Apr 21.
  20. Petit V et al. Optimization of tumor xenograft dissociation for the profiling of cell surface markers and nutrient transporters. Lab Invest 2013 May;93(5):611–21.
  21. Twarock S et al. Synthesis of hyaluronan in oesophageal cancer cells is uncoupled from the prostaglandin-cAMP pathway. Br J Pharmacol 2009 May;157(2):234–43.
  22. Asplund A et al. Expression profiling of microdissected cell populations selected from basal cells in normal epidermis and basal cell carcinoma. Br J Dermatol 2008 Mar;158(3):527–38.
  23. Teleki I et al. The potential prognostic value of connexin 26 and 46 expression in neoadjuvant-treated breast cancer. BMC Cancer 1350. Epub 2013/02/02.
  24. Kiflemariam S et al. Scalable in situ hybridization on tissue arrays for validation of novel cancer and tissue-specific biomarkers. PLoS One 2012;7(3):e32927.
  25. Nodin B et al. Discovery of dachshund 2 protein as a novel biomarker of poor prognosis in epithelial ovarian cancer. J Ovarian Res 2012 Jan 27;5(1):6.
  26. Sircoulomb F et al. ZNF703 gene amplification at 8p12 specifies luminal B breast cancer. EMBO Mol Med 2011 Mar; 3(3):153–166. Epub 2011 Feb 15.
  27. Cawthorn TR et al.Proteomic Analyses Reveal High Expression of Decorin and Endoplasmin (HSP90B1) Are Associated with Breast Cancer Metastasis and Decreased Survival. PLoS One 2012;7(2):e30992.
  28. Henke A et al. Stromal Expression of Decorin, Semaphorin6D, SPARC, Sprouty1 and Tsukushi in Developing Prostate and Decreased Levels of Decorin in Prostate Cancer. PLoS One 7(8):e42516. Epub 2012 Aug 3.
  29. Hudson EP et al. Multiplex epitope mapping using bacterial surface display reveals both linear and conformational epitopes. Sci Rep 2012;2:706.
  30. Arabi A et al. Proteomic screen reveals Fbw7 as a modulator of the NF-κB pathway. Nat Commun 2012;3:976.
  31. Ito A et al. Novel application for pseudopodia proteomics using excimer laser ablation and two-dimensional difference gel electrophoresis. Lab Invest 2012 Sep;92(9):1374–85.
  32. Holland DG et al. ZNF703 is a common Luminal B breast cancer oncogene that differentially regulates luminal and basal progenitors in human mammary epithelium. EMBO Mol Med 2011 Mar;3(3):167–80.
  33. Mulder J et al. Tissue profiling of the mammalian central nervous system using human antibody-based proteomics. Mol Cell Proteomics 2009 Jul;8(7):1612–22.
  34. Su D et al. Role of ERRF, a Novel ER-Related Nuclear Factor, in the Growth Control of ER-Positive Human Breast Cancer Cells. Am J Pathol 2012 Mar; 180(3):1189–1201.
  35. Shubbar E et al. High levels of γ-glutamyl hydrolase (GGH) are associated with poor prognosis and unfavorable clinical outcomes in invasive breast cancer. BMC Cancer 2013 Feb 1;13:47.
  36. Liao YC et al. Overexpressed hPTTG1 promotes breast cancer cell invasion and metastasis by regulating GEF-H1/RhoA signalling. Oncogene 2012 Jun 21;31(25):3086–97
  37. Cheng IK et al. GEF-H1 over-expression in hepatocellular carcinoma promotes cell motility via activation of RhoA signalling. Pathol 2012 Jul 30.
  38. Elkabets M et al. Human tumors instigate granulin-expressing hematopoietic cells that promote malignancy by activating stromal fibroblasts in mice. J Clin Invest 2011 Feb 1;121(2):784–99.
  39. Zibert JR et al. Halting angiogenesis by non-viral somatic gene therapy alleviates psoriasis and murine psoriasiform skin lesions. J Clin Invest 2011 Jan 4;121(1):410–21.
  40. Smyth LG et al. Carbonic anhydrase IX expression in prostate cancer. Prostate Cancer and Prostatic Diseases 2009 Dec;13(2):178–181.
  41. Paatero I et al. Interaction with ErbB4 promotes hypoxia-inducible factor-1α signaling. J Biol Chem 2012 Mar 23;287(13):9659–71.
  42. Zbytek B et al. Putative role of HIF transcriptional activity in melanocytes and melanoma biology. Dermatoendocrinol 2013 Apr 1; 5(2):239–251. Epub 2013/04/01.
  43. Hu Z et al. The expression level of HJURP has an independent prognostic impact and predicts the sensitivity to radiotherapy in breast cancer. Breast Cancer Res 2010;12(2):R18.
  44. Shuaib M et al. HJURP binds CENP-A via a highly conserved N-terminal domain and mediates its deposition at centromeres. Proc Natl Acad Sci U S A 2010 Jan 26;107(4):1349–54.
  45. de Tayrac M et al. Prognostic Significance of EDN/RB, HJURP, p60/CAF-1 and PDLI4, Four New Markers in High-Grade Gliomas. PLoS One 2013 Sep 11;8(9):e73332. 46. Bjarnadottir O et al. Targeting HMG-CoA reductase with statins in a window-ofopportunity breast cancer trial. Breast Cancer Res Treat 2013 Apr;138(2):499–508.
  46. Bjarnadottir O et al. Targeting HMG-CoA reductase with statins in a window-ofopportunity breast cancer trial. Breast Cancer Res Treat 2013 Apr;138(2):499–508.
  47. Jaraj SJ et al. GAD1 is a biomarker for benign and malignant prostatic tissue. Scand J Urol Nephrol 2011 Feb;45(1):39–45.
  48. Liu H et al. Single-cell clones of liver cancer stem cells have the potential of differentiating into different types of tumor cells. Cell Death Dis 2013 Oct; 4(10):e857-. Epub 2013/10/17.
  49. Guo L et al. Stat3-coordinated Lin-28-let-7-HMGA2 and miR-200-ZEB1 circuits initiate and maintain oncostatin M-driven epithelial-mesenchymal transition. Oncogene 2013 Nov 7;32(45):5272–82.
  50. Possemato R et al. Functional genomics reveal that the serine synthesis pathway is essential in breast cancer. Nature 2011 Aug 18;476(7360):346–50.
  51. Maddocks OD et al. Serine starvation induces stress and p53-dependent metabolic remodelling in cancer cells. Nature 2013 Jan 24;493(7433):542–6.
  52. Nilsson LM et al. Mouse genetics suggests cell-context dependency for Myc-regulated metabolic enzymes during tumorigenesis. PLoS Genet 2012;8(3):e1002573.
  53. Salem AF et al. Mitochondrial biogenesis in epithelial cancer cells promotes breast cancer tumor growth and confers autophagy resistance. Cell Cycle 2012 Nov 15; 11(22):4174–4180.
  54. Wagoner MP et al. The transcription factor REST is lost in aggressive breast cancer. PLoS Genet 2010 Jun 10;6(6):e1000979.
  55. Prada I et al. REST/NRSF governs the expression of dense-core vesicle gliosecretion in astrocytes. J Cell Biol 2011 May 2;193(3):537–49.
  56. Jögi A et al. Nuclear expression of the RNA-binding protein RBM3 is associated with an improved clinical outcome in breast cancer. Mod Pathol 2009 Dec;22(12):1564–74.
  57. Hjelm B et al. High nuclear RBM3 expression is associated with an improved prognosis in colorectal cancer. Proteomics Clin Appl 2011 Dec;5(11-12):624–35.
  58. Ehlén A et al. Expression of the RNA-binding protein RBM3 is associated with a favourable prognosis and cisplatin sensitivity in epithelial ovarian cancer. J Transl Med 2010 Aug 20;8:78.
  59. Jonsson L et al. High RBM3 expression in prostate cancer independently predicts a reduced risk of biochemical recurrence and disease progression. Diagn Pathol 2011 Sep 28;6:9.
  60. Nodin B et al. High MCM3 expression is an independent biomarker of poor prognosis and correlates with reduced RBM3 expression in a prospective cohort of malignant melanoma. Diagn Pathol 782. Epub 2012 Jul 17.
  61. Jonsson L et al. Low RBM3 protein expression correlates with tumour progression and poor prognosis in malignant melanoma: an analysis of 215 cases from the Malmö Diet and Cancer Study. J Transl Med 2011 Jul 21;9:114.
  62. Ehlén Å et al. RBM3-regulated genes promote DNA integrity and affect clinical outcome in epithelial ovarian cancer. Transl Oncol 2011 Aug;4(4):212–21.
  63. Hjelm B et al. High nuclear RBM3 expression is associated with an improved prognosis in colorectal cancer. Proteomics Clin Appl 2011 Dec;5(11-12):624–35.
  64. Boman K et al. Decreased expression of RNA-binding motif protein 3 correlates with tumour progression and poor prognosis in urothelial bladder cancer. BMC Urol 2013 Apr 8;13:17.
  65. Nodin B et al. High MCM3 expression is an independent biomarker of poor prognosis and correlates with reduced RBM3 expression in a prospective cohort of malignant melanoma. Diagn Pathol 2012 Jul 17;7:82.
  66. Telikicherla D et al. Overexpression of ribosome binding protein 1 (RRBP1) in breast cancer. Clin Proteomics 9(1):7. Epub 2012 Jun 18.
  67. Iwanaga R et al. Expression of Six1 in luminal breast cancers predicts poor prognosis and promotes increases in tumor initiating cells by activation of extracellular signalregulated kinase and transforming growth factor-beta signaling pathways. Breast Cancer Res 2012 Jul 5;14(4):R100.
  68. Smith AL et al. The miR-106b-25 cluster targets Smad7, activates TGF-β signaling, and induces EMT and tumor initiating cell characteristics downstream of Six1 in human breast cancer. Oncogene 2012 Jan 30.
  69. Wan F et al. Gene expression changes during HPV-mediated carcinogenesis: a comparison between an in vitro cell model and cervical cancer. Int J Cancer 2008 Jul 1;123(1):32–40.
  70. McCoy EL et al. Six1 expands the mouse mammary epithelial stem/progenitor cell pool and induces mammary tumors that undergo epithelial-mesenchymal transition. J Clin Invest 2009 Sep;119(9):2663–77.
  71. Micalizzi DS et al. The Six1 homeoprotein induces human mammary carcinoma cells to undergo epithelial-mesenchymal transition and metastasis in mice through increasing TGF-beta signaling. J Clin Invest 2009 Sep;119(9):2678–90.
  72. Farabaugh et al. Eya2 is required to mediate the pro-metastatic functions of Six1 via the induction of TGF-β signaling, epithelial-mesenchymal transition, and cancer stem cell properties. Oncogene 2012 Feb 2;31(5):552–62.
  73. Ono H et al. SIX1 promotes epithelial-mesenchymal transition in colorectal cancer through ZEB1 activation. Oncogene 2012 Nov 22;31(47):4923–34.
  74. Le Grand F et al. Six1 regulates stem cell repair potential and self-renewal during skeletal muscle regeneration. J Cell Biol 2012 Sep 3; 198(5):815–832.
  75. Sernbo S et al. Nuclear T-STAR Protein Expression Correlates with HER2 Status, Hormone Receptor Negativity and Prolonged Recurrence Free Survival in Primary Breast Cancer and Decreased Cancer Cell Growth In Vitro. PLoS One 8(7):e70596. Epub 2013/07/29.
  76. Ek S et al. From gene expression analysis to tissue microarrays: a rational approach to identify therapeutic and diagnostic targets in lymphoid malignancies. Mol Cell Proteomics 2006 Jun;5(6):1072–81.
  77. Schenke-Layland K et al. Cardiomyopathy is associated with structural remodelling of heart valve extracellular matrix. Eur Heart J 2009 Sep;30(18):2254–65.
  78. Ghosh Z et al. Dissecting the Oncogenic Potential of Human Embryonic and Induced Pluripotent Stem Cell Derivatives. Cancer Res 2011 Jul 15; 71(14):5030–5039. Epub 2011 Jun 6.
  79. Edlund K et al. CD99 is a novel prognostic stromal marker in non-small cell lung cancer. Int J Cancer 2012 Nov 15;131(10):2264–73.
  80. Pontén F et al. The Human Protein Atlas--a tool for pathology. J Pathol 2008 Dec;216(4):387–93.
  81. Wu CC et al. Candidate serological biomarkers for cancer identified from the secretomes of 23 cancer cell lines and the human protein atlas. Mol Cell Proteomics 2010 Jun;9(6):1100–17.
  82. Davidson B et al. Gene expression signatures differentiate ovarian/peritoneal serous carcinoma from breast carcinoma in effusions. J Cell Mol Med 2011 Mar;15(3):535–44.
  83. Dziegiel P et al. Ceramide galactosyltransferase (UGT8) is a molecular marker of breast cancer malignancy and lung metastases. Br J Cancer 2010 Aug 10;103(4):524–31.
  84. de Kruijf EM et al. NKG2D ligand tumor expression and association with clinical outcome in early breast cancer patients: an observational study. BMC Cancer 1224. Epub 2012 Jan 18.

Antibodies Against Gene Products in MammaPrint, Oncotype, EndoPredict and uPA Tests

This section presents antibodies in Prestige Antibody product catalog against gene products included in the diagnostic MammaPrint, EndoPredict, Oncotype and uPA tests. MammaPrint is a gene expression profile test based on the Amsterdam 70-gene breast cancer gene signature marketed by Agendia. It is a test to assess the risk that a breast tumor will metastasize to other parts of the body. MammaPrint aims at stratifying patients into “Low Risk” and “High Risk”. Oncotype DX (developed by Genomic Health) is the most frequently used gene expression profile in clinical practice in the United States analyzing a panel of 21 genes within a tumor to determine a Recurrence Score.

BIRC5/Survivin

IHC staining of human lung tissue using the Anti-CD68 antibody (HPA048982) shows strong cytoplasmic positivity in macrophages and in hematopoietic tissues, such as spleen.

CD68/Macrosialin

IHC staining of human lung tissue using the Anti-CD68 antibody (HPA048982) shows strong cytoplasmic positivity in macrophages and in hematopoietic tissues, such as spleen.

DTL

IHC staining of human bone marrow using the Anti-DTL antibody (HPA028016) shows strong nuclear positivity in bone marrow poietic cells. By ICC-IF, staining of nucleus in U-251 MG cells is detected.

GSTM5

The Anti-GSTM5 antibody (HPA048652) shows cytoplasmic positivity in glandular cells in human rectum by IHC and in WB, the antibody detects a band of predicted size in cell lysates of RT-4, U-251 MG, as well as in liver tissue lysate.
 

Product No. Target Protein Product Description Validated Applications
HPA002636 AURKA/STK15 Anti-AURKA IHC,WB
HPA012582 AZGP1 Anti-AZGP1 IHC,WB
HPA018121 BAG1 Anti-BAG1 IHC,ICC-IF
HPA002830 BIRC5/Survivin Anti-BIRC5 IHC,WB,ICC-IF
HPA048982 CD68/Macrosialin Anti-CD68 IHC
HPA005565 CDCA7 Anti-CDCA71,2 IHC,WB,ICC-IF
HPA006871 CMC2/C16orf61 Anti-CMC2 IHC
HPA044280 DHCR7 Anti-DHCR7 IHC,WB
HPA018670 DHX58/LGP2 Anti-DHX58 IHC,WB,ICC-IF
HPA019570 DHX58/LGP2 Anti-DHX58 IHC
HPA032152 DIAPH3 Anti-DIAPH3 IHC,WB*
HPA028016 DTL Anti-DTL3 IHC,WB,ICC-IF
HPA022130 ECI2/PECI Anti-ECI2 IHC,WB,ICC-IF
HPA022129 EGLN1/PHD2 Anti-EGLN14 IHC,ICC-IF
HPA036660 ESM1 Anti-ESM1 IHC,WB
HPA000449 Estrogen receptor Anti-ESR1 IHC,WB
HPA000450 Estrogen receptor Anti-ESR1 IHC,WB
HPA044394 Exostosin-1 Anti-EXT1 IHC,WB
HPA018795 FGF18 Anti-FGF18 IHC,WB,ICC-IF
HPA050682 GMPS Anti-GMPS IHC
HPA003011 GNAZ Anti-GNAZ IHC,WB
HPA017346 GPR126/VIGR Anti-GPR126 IHC
HPA047250 GPR180 Anti-GPR180 IHC,ICC-IF
HPA035190 GSTM3 Anti-GSTM3 IHC,WB
HPA048652 GSTM5/GSTM1 Anti-GSTM5 IHC,WB
HPA001383 HER2/ERBB2 Anti-ERBB2 IHC,WB
AMAb90627 HER2/ERBB2 Anti-HER2 IHC,WB
HPA051179 HRASLS Anti-HRASLS IHC
HPA010558 IL6ST/GP130 Anti-IL6ST5 IHC
HPA012114 JHDM1D/KDM7A Anti-JHDM1D IHC,ICC-IF

* WB both in human and rodent samples

  1. Gill RM et al. The MYC-Associated Protein CDCA7 Is Phosphorylated by AKT To Regulate MYC-Dependent Apoptosis and Transformation. Mol Cell Biol 2013 Feb; 33(3):498–513.
  2. Shubbar E et al. Elevated cyclin B2 expression in invasive breast carcinoma is associated with unfavorable clinical outcome. BMC Cancer 131. Epub 2013/01/02.
  3. Karaayvaz M et al. Prognostic significance of miR-215 in colon cancer. Clin Colorectal Cancer 2011 Dec;10(4):340–7.
  4. Bozóky B et al. Novel signatures of cancer-associated fibroblasts. Int J Cancer 2013 Jan 15.
  5. Rognum IJ et al. Interleukin-6 and the serotonergic system of the medulla oblongata in the sudden infant death syndrome. Acta Neuropathol 2009 Oct;118(4):519–3. S Alterations in Breast Cancer from Women Treated with Neoadjuvant Chemotherapy. PLoS One 2013;8(3):e60576.

MMP9

IHC staining of human lung tissue using the Anti-MMP9 antibody (HPA001238) shows strong nuclear positivity in macrophages and in bone marrow poietic cells in bone marrow tissue.

Monclonal Anti-MMP9 antibodies show strong cytoplasmic positivity in a subset of lymphoid cells in duodenum (AMAb90805) and in human tonsil tissue (AMAb90804).

LYRIC/MTDH

IHC staining using the Anti-MTDH antibody (HPA010932) shows strong cytoplasmic positivity in neuronal cells in human cerebral cortex tissue. In ICC-IF in A-431 cell line, the antibody stains endoplasmic reticulum.

IHC staining using the monclonal Anti-MTDH antibody (AMAb90762) shows strong cytoplasmic reactivity in tumor cells from breast and colorectal cancer samples.

P5C Dehydrogenase/ALDH4A1

IHC staining using the Anti- ALDH4A1 antibody (HPA006401) shows strong cytoplasmic positivity with granular pattern in human kidney and liver tissues.
 

Product No. Target Protein Product Description Validated Applications
HPA000451 Ki67/MKI67 Anti-MKI671 IHC,ICC-IF
HPA001164 KI67/MKI67 Anti-MKI672 IHC,ICC-IF
AMAb90870 KI67/MKI67 Anti-MKI67 IHC
HPA030241 LIN9 Anti-LIN9 IHC,ICC-IF
HPA012501 LPCAT/AYTL2 Anti-LPCAT1 IHC,WB
HPA022268 LPCAT/AYTL2 Anti-LPCAT13 IHC,WB,ICC-IF
HPA015104 LYRIC Anti-MTDH4 IHC,WB,ICC-IF
HPA010932 LYRIC Anti-MTDH5 IHC,WB*,ICC-IF
AMAb90762 LYRIC Anti-MTDH IHC,WB
AMAb90763 LYRIC Anti-MTDH IHC,WB
HPA013949 Matrix Gla protein Anti-MGP6 IHC
HPA004818 MCM6 Anti-MCM6 IHC,WB*,ICC-IF
HPA017214 MELK/PK38 Anti-MELK IHC,ICC-IF
HPA001238 MMP9 Anti-MMP9 IHC,WB,ICC-IF
AMAb90804 MMP9 Anti-MMP9 IHC,WB
AMAb90805 MMP9 Anti-MMP9 IHC,WB
AMAb90806 MMP9 Anti-MMP9 IHC
HPA017418 MS4A7 Anti-MS4A7 IHC,WB
HPA030530 MYBL2 Anti-MYBL2 IHC,WB
HPA025926 Neuromedin-U Anti-NMU IHC,WB
HPA042904 NUSAP1 Anti-NUSAP1 IHC,ICC-IF
HPA006401 P5C dehydrogenase Anti-ALDH4A1 IHC,WB

* WB both in human and rodent samples

  1. Pohler E et al. Haploinsufficiency for AAGAB causes clinically heterogeneous forms of punctate palmoplantar keratoderma. Nat Genet 2012 Oct 14;44(11):1272–6.
  2. Roca H et al. IL-4 induces proliferation in prostate cancer PC3 cells under nutrient-depletion stress through the activation of the JNK-pathway and survivin upregulation. J Cell Biochem 2012 May; 113(5):1569–1580.
  3. Friedman JS et al. Loss of lysophosphatidylcholine acyltransferase 1 leads to photoreceptor degeneration in rd11 mice. Proc Natl Acad Sci U S A 2010 Aug 31;107(35):15523–8.
  4. Nohata N et al. Tumor suppressive microRNA-375 regulates oncogene AEG-1/ MTDH in head and neck squamous cell carcinoma (HNSCC). J Hum Genet 2011 Aug;56(8):595–601.
  5. Liu B et al. Astrocyte elevated gene-1 regulates osteosarcoma cell invasion and chemoresistance via endothelin-1/endothelin A receptor signaling. Oncol Lett 2013 Feb;5(2):505–510.
  6. Lorenzen JM et al. Fetuin, matrix-Gla protein and osteopontin in calcification of renal allografts. PLoS One 2012;7(12):e52039.

PITRM1/MP1

The Anti- PITRM1 antibody (HPA006753) shows strong cytoplasmic positivity in myocytes in human heart muscle using IHC. ICC-IF staining of human cell line U-251 MG shows positivity in mitochondria.

PRC1

IHC staining of human testis tissue using the Anti-PRC1 antibody (HPA034521) shows strong nuclear positivity in cells of seminiferus ducts. ICC-IF shows staining of nucleus, plasma membrane and microtubules in A-431 cells.

SCOT/OXCT1

IHC staining of human heart muscle and kidney by Anti-OXCT1 antibody (HPA012047) shows strong cytoplasmic positivity in myocytes and cells in tubules, respectively. ICC-IF shows staining of mitochondria in A431 cells.
 

Product No. Target Protein Product Description Validated Applications
HPA006753 PITRM1/MP1 Anti-PITRM1 IHC,WB,ICC-IF
HPA006754 PITRM1/MP1 Anti-PITRM1 IHC,WB*,ICC-IF
HPA008719 PLAU/UPA Anti-PLAU IHC,WB
HPA034521 PRC1 Anti-PRC1 IHC,ICC-IF
HPA004751 Progesteron receptor Anti-PGR1 IHC,ICC-IF
HPA008428 Progesteron receptor Anti-PGR IHC
HPA017176 Progesteron receptor Anti-PGR IHC
HPA012716 QSOX2/QSCN6L1 Anti-QSOX2 IHC,WB,ICC-IF
HPA039890 RBBP8 Anti-RBBP8 IHC, WB
HPA029970 RECQL5 Anti-RECQL5 IHC,ICC-IF
HPA029971 RECQL5 Anti-RECQL5 IHC,WB,ICC-IF
HPA044428 RTN4RL1/NgR3 Anti-RTN4RL1 IHC
HPA023726 RUNDC1 Anti-RUNDC1 IHC,WB,ICC-IF
HPA006353 SCUBE2/CEGP1 Anti-SCUBE2 IHC,ICC-IF
HPA029871 SCUBE2/CEGP1 Anti-SCUBE2 IHC
HPA012047 SCOT/OXCT1 Anti-OXCT12 IHC,WB*,ICC-IF
HPA050039 SERPINE1/PAI1 Anti-SERPINE1 IHC
HPA006539 SLC2A3/GLUT3 Anti-SLC2A3 IHC
HPA045372 Stanniocalcin-2 Anti-STC2 IHC, WB, IF
HPA015820 STK32B Anti-STK32B IHC,ICC-IF
HPA027923 TGFB3 Anti-TGFB3 IHC,WB
HPA045213 TMEM74B/C20orf46 Anti-TMEM74B IHC
HPA031347 TSPYL5 Anti-TSPYL5 IHC
HPA005908 UCHL5 Anti-UCHL5 IHC,ICC-IF
HPA014290 VEGFR-1 Anti-FLT1 IHC,ICC-IF
AMAb90703 VEGFR-1 Anti-FLT1 IHC
AMAb90704 VEGFR-1 Anti-FLT1 IHC,WB
HPA007121 WISP1 Anti-WISP1 IHC,ICC-IF

* WB both in human and rodent samples

  1. Pereira CB et al. Prognostic and Predictive Significance of MYC and KRAS Alterations in Breast Cancer from Women Treated with Neoadjuvant Chemotherapy. PLoS One 2013;8(3):e60576.
  2. Chang HT et al. Ketolytic and glycolytic enzymatic expression profiles in malignant gliomas: implication for ketogenic diet therapy. Nutr Metab (Lond) 1047. Epub 2013/07/05.
  3. Zibert JR et al. Halting angiogenesis by non-viral somatic gene therapy alleviates psoriasis and murine psoriasiform skin lesions. J Clin Invest 2011 Jan 4;121(1):410–21.

Antibodies Identified in the Human Protein Atlas

Showing Differential IHC Staining Patterns in Breast Cancer Samples

IHC analysis using Anti-KLHL26 antibody (HPA023074) shows a varying membranous/cytoplasmic staining pattern in breast tumor samples from different patients.

The Anti-ACSF2 (HPA024693) antibody shows granular cytoplasmic positivity in breast tumor cells from different patients varying from strong to negative.

The Anti-GCM1 (HPA011343) antibody shows membranous positivity in breast tumor cells while normal breast tissue is negative.

The Anti-AGR3 (HPA053942) antibody shows strong cytoplasmic positivity in 11/12 breast cancer patients, while 1 patient is completely negative.
 

Product No. Product Description Validated Applications
HPA037918 Anti-AAMDC IHC,WB
HPA037919 Anti-AAMDC IHC
HPA040312 Anti-ABCG4 IHC,ICC-IF
HPA007695 Anti-NIM1 IHC,WB,ICC-IF
HPA023993 Anti-AC145676.2 IHC,WB
HPA024693 Anti-ACSF2 IHC,WB
HPA042014 Anti-ADAMTS13 IHC,WB
HPA053942 Anti-AGR3 IHC
HPA020522 Anti-AIF1L IHC,WB
HPA006171 Anti-AJUBA IHC, WB
HPA046271 Anti-ALDH1A3 IHC,WB
HPA013758 Anti-ANKRD46 IHC,WB
HPA004341 Anti-ASB6 IHC,WB,ICC-IF
HPA005935 Anti-ATF6 IHC
HPA008147 Anti-ATP6V1B2 IHC,WB*,ICC-IF
HPA046678 Anti-AVPR2 IHC
HPA020274 Anti-BCL9 IHC
HPA038478 Anti-BTG4 IHC
HPA026810 Anti-ADIRF IHC,WB
HPA007968 Anti-C10orf54 IHC,WB,ICC-IF
HPA039713 Anti-C12orf76 IHC,WB
HPA008959 Anti-C17orf85 IHC
HPA045811 Anti-C1ORF195 IHC,ICC-IF
HPA051143 Anti-C2orf68 IHC
HPA037889 Anti-SIMC1 IHC,WB
HPA021480 Anti-CAPN8 IHC,WB
HPA023457 Anti-CCDC144NL IHC,WB
HPA027185 Anti-CCDC170 IHC,WB
HPA027121 Anti-CCDC170 IHC,WB
HPA002637 Anti-CDK6 IHC,WB*,ICC-IF
HPA014361 Anti-CLDN3 IHC
HPA041132 Anti-CPNE2 IHC,WB
HPA004135 Anti-CRABP2 IHC,WB,ICC-IF
HPA015077 Anti-CTNND2 IHC
HPA006128 Anti-CXorf67 IHC,WB
HPA017661 Anti-CYP4X1 IHC
HPA012672 Anti-DACH1 IHC,ICC-IF
HPA051589 Anti-DBF4 IHC
HPA050246 Anti-DCHS1 IHC
HPA015655 Anti-DCLK1 IHC
HPA047631 Anti-DECR2 IHC
HPA046708 Anti-DOM3Z IHC
HPA018221 Anti-DUSP26 IHC,WB
HPA006465 Anti-ECD IHC,WB,ICC-IF
HPA049331 Anti-EFHD1 IHC,WB,ICC-IF
HPA007397 Anti-EPHA6 IHC
HPA030879 Anti-FAM101B IHC
HPA009410 Anti-FAM189A1 IHC
HPA008707 Anti-FKBP7 IHC,WB*,ICC-IF
HPA004937 Anti-FMN2 IHC
HPA052324 Anti-G6PC IHC
HPA044371 Anti-GABRD IHC,WB
HPA027463 Anti-GAK IHC,ICC-IF
HPA011343 Anti-GCM1 IHC
HPA039916 Anti-GLB1L3 IHC
HPA002318 Anti-GLDC IHC,WB*
HPA039501 Anti-GLYATL1 IHC,WB
HPA007990 Anti-GTF3A IHC,ICC-IF
HPA007611 Anti-HIPK2 IHC,ICC-IF
HPA036913 Anti-HMGCS1 IHC,WB,ICC-IF
HPA027423 Anti-HMGCS2 IHC,WB
HPA027442 Anti-HMGCS2 IHC,WB
HPA004337 Anti-IFITM3 IHC,WB
HPA054669 Anti-IRX2 IHC,WB
HPA007931 Anti-ISYNA1 IHC
HPA008232 Anti-ISYNA1 IHC,WB
HPA008572 Anti-ITGA3 IHC
HPA005676 Anti-ITGBL1 IHC,WB
HPA000506 Anti-ITIH6 IHC
HPA051637 Anti-KLHL2 IHC
HPA023074 Anti-KLHL26 IHC,WB
HPA049550 Anti-KRT31 IHC
HPA040330 Anti-KRT32 IHC
HPA042482 Anti-KRTAP9-3 IHC
HPA012072 Anti-LASP11 IHC,WB*,ICC-IF
HPA008556 Anti-LGR6 IHC
HPA036706 Anti-LRRIQ4 IHC
HPA002820 Anti-MAGEB1 IHC
HPA039454 Anti-MANSC4 IHC,WB
HPA059457 Anti-MROH2B IHC
HPA017642 Anti-MRS2 IHC,WB
HPA005914 Anti-MSTO1 IHC
HPA049831 Anti-MTMR2 IHC
HPA005466 Anti-MYBBP1A IHC,WB,ICC-IF
HPA024338 Anti-NAPEPLD IHC,WB,ICC-IF
HPA028136 Anti-NASP IHC,WB,ICC-IF
HPA006111 Anti-NFIA IHC,WB*,ICC-IF
HPA006873 Anti-NKAIN1 IHC
HPA007489 Anti-NPSR12 IHC,ICC-IF
HPA048760 Anti-OR2Z1 IHC
HPA015808 Anti-OR9K2 IHC
HPA024524 Anti-OTOP2 IHC
HPA048975 Anti-PDE4C IHC,WB
HPA051038 Anti-PEG10 IHC,ICC-IF
HPA003994 Anti-PHLDA2 IHC
HPA020200 Anti-PHLPP1 IHC
HPA012312 Anti-PHTF2 IHC
HPA045390 Anti-PKN3 IHC
HPA044690 Anti-PNMA5 IHC,ICC-IF
HPA051607 Anti-PPP1R35 IHC
HPA023923 Anti-PPR11 IHC,WB,ICC-IF
HPA048536 Anti-PVALB IHC
HPA019717 Anti-RAB313 IHC,WB*
HPA047820 Anti-RAC3 IHC,WB
HPA008752 Anti-RAD18 IHC,WB
HPA058061 Anti-REEP1 IHC
HPA005681 Anti-RIOK2 IHC,ICC-IF
HPA015733 Anti-RNF152 IHC,WB
HPA005985 Anti-RPS13 IHC
HPA006462 Anti-S100A1 IHC,WB,ICC-IF
HPA019592 Anti-S100A13 IHC,WB*
HPA027613 Anti-S100A14 IHC,ICC-IF
HPA006997 Anti-S100A7 IHC,ICC-IF
HPA013321 Anti-SGK196 IHC,WB,ICC-IF
HPA051248 Anti-SH3BGRL IHC,WB
HPA037690 Anti-SHROOM1 IHC
HPA005911 Anti-SLC16A7 IHC,WB
HPA042377 Anti-SLC39A6 IHC,WB
HPA023748 Anti-SPAG1 IHC,ICC-IF
HPA018038 Anti-SQLE IHC,WB
HPA011173 Anti-SRPRB IHC,WB
HPA039789 Anti-SSSCA1 IHC,WB*,ICC-IF
HPA049106 Anti-STAG3 IHC,WB,ICC-IF
HPA042583 Anti-STARD6 IHC,IF
HPA001467 Anti-STX74 IHC,WB*,ICC-IF
HPA005781 Anti-TACC3 IHC,WB
HPA007066 Anti-TAPBP IHC
HPA000262 Anti-TBC1D9 IHC,ICC-IF
HPA046156 Anti-TCTE3 IHC
HPA017019 Anti-TGFBI IHC
HPA051855 Anti-TMEM110-MUSTN1 IHC
HPA016579 Anti-TMEM222 IHC,ICC-IF
HPA046658 Anti-TMEM47 IHC
HPA018216 Anti-TMEM68 IHC,ICC-IF
HPA005487 Anti-TPX2 IHC,WB,ICC-IF
HPA003054 Anti-TTLL12 IHC,WB
HPA023605 Anti-UBE20 IHC,WB*
HPA042302 Anti-WFDC2 IHC
HPA050514 Anti-WNT3A IHC
HPA006811 Anti-ZBTB7B IHC,WB*,ICC-IF
HPA009637 Anti-ZKSCAN3 IHC,ICC-IF
HPA007023 Anti-ZNF131 IHC
HPA049770 Anti-ZNF627 IHC,WB
HPA039116 Anti-ZNF662 IHC,WB

* WB both in human and rodent samples

  1. Ngan E et al. A complex containing LPP and α-Actinin mediates TGF β-induced migration and invasion of ErbB2-expressing breast cancer cells. J Cell Sci 2013 May 1; 126(0 9):1981–1991. Epub 2013/02/27.
  2. Camilleri M et al. Neuropeptide S receptor induces neuropeptide expression and associates with intermediate phenotypes of functional gastrointestinal disorders. Gastroenterology 2010 Jan;138(1):98–107.e4.
  3. Bozóky B et al. Novel signatures of cancer-associated fibroblasts. Int J Cancer 2013 Jan 15.
  4. Strömberg S et al. Selective expression of Syntaxin-7 protein in benign melanocytes and malignant melanoma. J Proteome Res 2009 Apr;8(4):1639–46.

Finding Cancer Biomarkers

Breast Cancer

Breast cancer is the second most common cancer and by far the most frequent cancer among women. The incidence of breast cancer is increasing steadily, but without a corresponding increase in mortality. If detected at an early stage, the prognosis is relatively good for a patient living in a developed country, with a general five-year survival rate of approximately 85%.

Breast Cancer and Treatment

Cancer, though often denoted as a singular disease, is truly a multitude of diseases. This understanding has evolved over the years, but many patients are not receiving optimal treatment for their disease. For cancer patients to receive a more individualized treatment, there is still a need for new and better ways to stratify patients. The classical prognostic factors such as stage and grade of the tumor are insufficient for a correct estimation of patient prognosis. Additional information from cancer biomarkers promise to substantially improve this estimation, ultimately leading to a more individualized treatment, thus avoiding both under and overtreatment of patients.

The primary curative treatment for breast cancer patients is surgery, often in combination with adjuvant therapy. However, adjuvant therapy is associated with substantial costs and sometimes severe side effects, and physicians have identified reduction of overtreatment as the major clinical need in breast cancer treatment today. Thus, the stratification of patients into different prognostic categories is of great importance as it may aid physicians in selecting the most appropriate treatment for a given patient.

The majority of breast cancers are hormone receptor responsive, i.e., express the estrogen receptor (ER) and/or the progesteron receptor (PR). Patients with tumors expressing these receptors may receive adjuvant endocrine treatment, such as tamoxifen.

Breast cancers may also express the HER2 protein (human epidermal growth factor receptor 2), and patients with tumors expressing this protein may receive adjuvant therapy with trastuzumab.

Adjuvant treatment may also consist of chemotherapy or radiation therapy.

RBM3

The RNA-binding motif protein 3 (RBM3) is an RNA- and DNAbinding protein, whose function has not been fully elucidated. It has been shown that the protein is expressed as an early event in mild hypothermia, and also in other conditions relating to cellular stress, such as glucose deprivation and hypoxia1. During stress, RBM3 is thought to protect the cells by aiding in maintenance of protein synthesis needed for survival1. Recently, it has also been shown that RBM3 attenuates stem cell-like properties in prostate cancer cells2.

RBM3 was identified via the Human Protein Atlas (HPA) as a potential oncology biomarker through the differential expression pattern present in several cancers investigated as part of the HPA project (proteinatlas.org)3,4.

The IHC analysis using the Anti-RBM3 antibody HPA003624 showed a weak expression pattern in normal breast tissue, but a stratified pattern in breast cancer tissue (Figure 1). Researchers further investigated the expression in larger breast cancer cohorts and the expression of RBM3 was shown to be associated with a prolonged survival5.

  1. Ehlén Å (2011) PhD Thesis: The role of RNA-binding motif 3 in epithelial ovarian cancer: A biomarker discovery approach.
  2. Zeng Y et al. (2013) Stress response protein RBM3 attenuates the stem-like properties of prostate cancer cells by interfering with CD44 variant splicing. Cancer Res. May 10. [Epub ahead of print].
  3. Berglund L et al. (2008) A gene-centric human protein atlas for expression profiles based on antibodies. Molecular & Cellular Proteomics 7:2019–2027.
  4. Uhlén M et al. (2010) Towards a knowledge-based Human Protein Atlas. Nat Biotechnol 28(12):1248–50.
  5. Jögi A et al. (2009) Nuclear expression of the RNA-binding protein RBM3 is associated with an improved clinical outcome in breast cancer. Mod Pathol. Dec;22(12):1564–74.

Figure 1. Immunohistochemical analysis using the Anti-RBM3 antibody (HPA003624) shows weak expression in normal breast tissue (A) and differential expression, varying from weak to strong in tumor breast samples (B, C).
 

RBM3 as a Prognostic Biomarker in Breast Cancer

After identification of RBM3 as a potential prognostic biomarker, researchers further investigated the RBM3 protein expression in larger breast cancer cohorts1. In a cohort of 500 premenopausal women with stage II invasive breast cancer, RBM3 expression was found to be associated with small, low-grade, estrogen receptor (ER)-positive tumors. When analyzing the subset of ER-positive patients, RBM3 was an independent predictor of recurrence free survival (RFS). As shown in Figure 2, patients with tumors expressing high levels of the RBM3 protein have an improved survival compared to patients with tumors expressing low levels of RBM3.

RBM3 protein expression has further been analyzed in many different patient cohorts from various forms of cancer. Levels of RBM3 expression was found to have a significant connection to patient survival in breast1, colon2, ovarian3,4, testicular, urothelial5, and prostate6 cancer as well as in malignant melanoma7.

In conclusion, RBM3 is a marker of good prognosis in breast cancer as well as in several other cancers.

Figure 2. Kaplan-Meier (survival) analysis of recurrence free survival (RFS) according to RBM3 expression for ER-positive breast cancer patients. Patients were split into two groups based on high and low RBM3 expression.

 

RBM3 Antibodies

There are two Anti-RBM3 antibodies in Atlas Antibodies' product catalog; the Triple A Polyclonal HPA003624 and the PrecisA Monoclonal AMAb90655. The monoclonal Anti-RBM3 antibody AMAb90655 has shown excellent specificity in Western Blot analysis of human cell lines, and is routinely used for staining of formalin fixed paraffin embedded tissue in IHC (Figure 3).

Figure 3. Immunohistochemical analysis of RBM3 expression in breast cancer (left) and prostate cancer (right) using AMAb90655 shows nuclear positivity in tumor cells. The WB image shows an expected band of 17 kDa in human cell line RT4 lysate using AMAb90655.

  1. Jögi A et al. (2009) Nuclear expression of the RNA-binding protein RBM3 is associated with an improved clinical outcome in breast cancer. Mod Pathol. Dec;22(12):1564–74.
  2. Hjelm B et al. (2011) High nuclear RBM3 expression is associated with an improved prognosis in colorectal cancer. Proteomics Clin Appl. Dec;5(11–12):624–35.
  3. Ehlén Å et al (2010) Expression of the RNA-binding protein RBM3 is associated with a favourable prognosis and cisplatin sensitivity in epithelial ovarian cancer. J Transl Med. Aug 20; 8:78.
  4. Ehlén Å et al. (2011) RBM3-regulated genes promote DNA integrity and affect clinical outcome in epithelial ovarian cancer. Transl Oncol. Aug;4(4):212–21.
  5. Boman K et al. (2013) Decreased expression of RNA-binding motif protein 3 correlates with tumour progression and poor prognosis in urothelial bladder cancer. BMC Urol. 2013;13:17.
  6. Jonsson L et al. (2011) High RBM3 expression in prostate cancer independently predicts a reduced risk of biochemical recurrence and disease progression. Diagn Pathol. Sep 28;6:91.
  7. Jonsson L et al. (2011) Low RBM3 protein expression correlates with tumour progression and poor prognosis in malignant melanoma: an analysis of 215 cases from the Malmö Diet and Cancer Study. J Transl Med. Jul 21;9:114.

Granulin

Granulins are a family of secreted, glycosylated peptides that are cleaved from a single precursor protein. Cleavage of the signal peptide produces mature granulin which can be further cleaved into a variety of active peptides. These cleavage products are named granulin A, granulin B, granulin C, etc. Both the peptides and intact granulin protein regulate cell growth. Different members of the granulin protein family may act as inhibitors, stimulators, or have dual actions on cell growth. Granulin family members are important in normal development, wound healing, and tumorigenesis [provided by RefSeq, Jul 2008].

In a paper by Elkabets et al, the role of GRN expression in responding tumor instigation was investigated by studying recrution of GRNexpressing bone marrow cells into responding tumors in mice1. Certain tumors can foster the growth of other tumors or metastatic cells located at distant anatomical sites, which is referred to as tumor instigation. In this study, rigorously growing human breast carcinoma cells were implanted in mice and it was shown that these cells stimulated the outgrowth of otherwise poorly tumorigenic, indolent transformed cells. GRN was identified as the most upregulated gene in the instigating bone marrow cells. The GRN expressing cells induced resident fibroblasts to express genes that promoted malignant tumor progression. It was speculated whether anticancer therapies might involve targeting GRN, or the activated GRN expressing cells, and thereby disrupting these cell lines of communication that promote cancer progression.

By using the Anti-GRN antibody HPA028747 in the analysis of tumor tissues from a cohort of breast cancer patients, high GRN expression was shown to correlate with the most aggressive triple-negative, basal-like tumor subtype and reduced patient survival (Figure 4).

  1. Elkabets M et al. Human tumors instigate granulin-expressing hematopoietic cells that promote malignancy by activating stromal fibroblasts in mice. J Clin Invest 2011 Feb 1;121(2):784–99.

Figure 4. GRN expression was shown to correlate with aggressive tumor subtypes and reduced survival of breast cancer patients using antibody HPA028747. The diagram to the left shows percentage of tumors in each category (Triple-Negative [TN]/ basal or nonbasal) that show high GRN positivity and the Kaplan-Meier analysis to the right shows correlation between GRN-positive (green) or GRN-negative (blue) expression and survival.

 

Granulin Antibodies

In Atlas Antibodies´ product catalog, there are two polyclonal Anti-GRN antibodies; HPA008763 and HPA028747.

IHC staining of human pancreas tissue using the Anti-GRN antibody (HPA008763) shows strong cytoplasmic positivity in exocrine glandular cells. ICC-IF shows positivity in vesicles in A-431 cells.

IHC analysis using the Anti-GRN antibody HPA028747 shows strong cytoplasmic positivity in normal duodenum tissue in glanduclar cells and vesicle positivity in U-251 MG cells.

 

Anillin

Anillin is an actin binding protein that is a subunit of microfilaments, one of the cytoskeleton components. Anillin is expressed in most cells and is involved in basic cell functions, e.g. motility, division and signaling. Studies of anillin expression have shown that it is over expressed in several human tumors.

Anillin as a Treatment Predictive Prognostic Biomarker in Breast Cancer

Anillin expression was analyzed in a patient cohort consisting of 467 samples from patients diagnosed with breast cancer, using the Anti-ANLN antibody HPA005680. Patients with tumors expressing high levels of anillin had a reduced recurrence free survival (RFS) compared to patients with tumors expressing low levels of anillin (Figure 5A). The same association between anillin expression and reduced survival could be seen when analyzing breast cancer specific survival (BCSS, Figure 5B). In a study by O´Leary et al, the prognostic impact of anillin was confirmed by Cox regression analysis. High anillin expression was associated with reduced BCSS and RFS in univariate- as well as in multivariate analysis, adjusted for tumor size and grade, age at diagnosis, nodal-, ER-, PR-, HER2-, and Ki67 status.

In conclusion, anillin is a marker for poor prognosis in breast cancer.

Figure 5. Kaplan-Meier (survival) analysis of recurrence free- (A) and breast cancer specific survival (B) according to aniliin expression for breast cancer patients. Patients were split into two groups based on high and low anillin expression.

  1. O´Leary PC et al. Systematic antibody generation and validation via tissue microarray technology leading to identification of a novel protein prognostic panel in breast cancer. BMC Cancer. 2013 Apr 2;13:175.

 

Anillin Antibodies

There are three Anti-ANLN antibodies in Atlas Antibodies product catalog; the PrecisA Monoclonals AMAb90660 and AMAb90662 and the Triple A Polyclonal HPA005680.

The Anti-ANLN antibody (HPA005680) shows strong nuclear positivity in cells in seminiferous ducts in human testis by IHC. In ICC-IF, nuclei (but not nucleoli) of A-431 cells stain positively and in WB, the antibody detects a band of predicted size in cell lysates of RT-4 and U-251 MG.

Anti-ANLN antibody AMAb90660 shows strong nuclear immunoreactivity in a subset of tumour cells in lung adenocarcinoma and a band of predicted size in human cell line U-251 MG.

AMAb90662 Anti-ANLN antibody shows strong nuclear immunoreactivity in a subset of tumor cells in colorectal cancer and a band of predicted size in human U-251 MG cells.