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MAB4190 Sigma-Aldrich

Anti-Ki-67 Antibody, clone Ki-S5

clone Ki-S5, Chemicon®, from mouse

  •  eCl@ss 32160702

  •  NACRES NA.41

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Properties

Related Categories Alphabetical Index, Antibodies, KIA-KIT, Primary Antibodies
clone   Ki-S5, monoclonal
biological source   mouse
application(s)   flow cytometry: suitable
  immunocytochemistry: suitable
  immunohistochemistry: suitable (paraffin)
  immunohistochemistry: suitable
  western blot: suitable
species reactivity   human
shipped in   ambient
packaging   antibody small pack of 25 μg
isotype   IgG1
Quality Level   100
antibody product type   primary antibodies
mfr. no.   Chemicon®
NCBI accession no.   NM_002417.3
UniProt accession no.   P46013

Description

General description

Ki67 antigen is the prototypic cell cycle related nuclear protein, expressed by proliferating cells in all phases of the active cell cycle (G1, S, G2 and M phase). It is absent in resting (G0) cells. Ki67 antibodies are useful in establishing the cell growing fraction in neoplasms (immunohistochemically quantified by determining the number of Ki67 positive cells among the total number of resting cells = Ki67 index). In neoplastic tissues the prognostic value is comparable to the tritiated thymidine labelling index. The correlation between low Ki67 index and histologically low grade tumours is strong. Ki67 is routinely used as a marker of cell cycling and proliferation. This specificity of Ki-S5 antibody for proliferating cells might make it a useful tool for determination of the proliferative fraction in tumors such as Non-Hodgkin′s lymphoma (Kreipe et al., 1993), mammary carcinomas (Mauri et al., 1984), ovarian tumors (Ballin et al., 1994) and prostate cancer.

Specificity

In Immunoprecipitation and Western blot experiments the Ki-S5 antibody recognizes a cell-cycle-associated protein of 345 kD and 395 kD (Kreipe et al., 1993) identical with the Ki-67 antigen. The antibody binds to a formalin-resistant epitope of the Ki-67 antigen. The immunoreactivity of Ki-S5 is confined to the nuclei of proliferating cells and no crossreactivity with cytoplasmic antigens of epithelial occurs even after antigen retrieval (Mauri et al., 1984). A comparison of immunohistochemical labeling of fresh and fixed tissue samples of NHL showed that identical results were obtained with Ki-67 and Ki-S5 (Kreipe et al., 1993). The Ki-67 antigen is preferentially expressed during late G1, S, G2 and M phase of the cell cycle, while resting, non-cycling cells (G0 phase) lack the Ki-67 antigen. In addition, constantly proliferating cells (e.g. cell lines) react positively to Ki-S5 during the entire cell-cycle. This specificity of Ki-S5 antibody for proliferating cells might make it a useful tool for determination of the proliferative fraction in tumors such as Non-Hodgkin′s lymphoma (Kreipe et al., 1993), mammary carcinomas (Mauri et al., 1984), ovarian tumors (Ballin et al., 1994) and prostate cancer.

Immunogen

Nuclear protein preparation from the human cell line U937.50.

Application

Anti-Ki-67 Antibody, clone Ki-S5 is a high quality Mouse Monoclonal Antibody for the detection of Ki-67 & has been validated in FC, ICC, IHC, IHC(P) & WB.

Immunocytochemistry:
A 5-10 μg/mL concentration of a previous lot was used in IC.

Immunohistochemistry:
A 5-10 μg/mL concentration of a previous lot was used in IH.

Immunohistochemistry:
5-10 µg/mL on formalin fixed paraffin tissue. Citrate buffer-microwave antigen retrieval

Flow Cytometry:
A previous lot of this antibody was used in FC.

Western blot:
1-10 µg/mL

Optimal working dilutions must be determined by end user.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Cell Cycle, DNA Replication & Repair

Target description

345 & 395 kDa

Physical form

Format: Purified

Mouse monoclonal IgG1 in buffer containing 0.02 M Phosphate buffer, 0.25 M NaCl, pH 7.6 with 0.1% sodium azide.

Protein A Purfied

Storage and Stability

Stable for 1 year at 2-8ºC from date of receipt.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Quality

Evaluated by Western Blot on A431 lysates.

Western Blot Analysis:
1:500 dilution of this antibody detected cell-cycle-associated protein of 345 kDa and 395 kDa on 10 μg of A431 lysates.

Analysis Note

Control
Tonsil tissue, A431 cell lysate

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Safety & Documentation

Safety Information

WGK Germany 
WGK 1
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable
Protocols & Articles

Articles

Antibody Basics

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Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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