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Analysis Note

ControlTonsil tissue, A431 cell lysate

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.

Research CategoryEpigenetics & Nuclear Function

Research Sub CategoryCell Cycle, DNA Replication & Repair

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.

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.

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.

Immunogen

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

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

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

Physical form

Protein A purified

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

Format: Purified

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.

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.

Storage and Stability

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

Target description

345 & 395 kDa