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MAB4190 Sigma-Aldrich

Anti-Ki-67 Antibody, clone Ki-S5

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.

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Properties

Related Categories Antibodies, Primary Antibodies More...
trade name   Chemicon
brand family   Chemicon
format   Purified
control   Tonsil tissue, A431 cell lysate
molecular weight   345 & 395 kDa
application(s)   Flow Cytometry
  Immunocytochemistry
  Immunohistochemistry
  Immunohistochemistry (Paraffin)
  Western Blotting
isotype   IgG1
clone   Monoclonal Antibody
  Ki-S5
purification method   Protein A Purfied
concentration   Please refer to the Certificate of Analysis for the lot-specific concentration.
host   Mouse
species reactivity   Human
NCBI accession no.   NM_002417.3
UniProt accession no.   P46013
gene symbol   MKI67(4288)
  KI-67
  KIA
material size   100 µg
shipped in   wet ice
storage conditions   Stable for 1 year at 2-8ºC from date of receipt.

Description

Background information

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.

Physical form

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

Application

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.

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.

Quality Assurance

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.

Usage Statement

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.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Articles

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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, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

Peer-Reviewed Papers
15

References

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