HCA117A recognises the sequence GFKELFQ, which can be found eight times on the human Ki67 protein. Ki-67 antigen is expressed in proliferating cells but not in quiescent cells. Expression of this antigen occurs preferentially during late G1, S, G2, and M phases of the cell cycle, while in cells in G0 phase the antigen cannot be detected. Consequently, Ki-67 antigen expression is used in tumor pathology to detect proliferating cells in neoplastic diseases. In cultured cells, Ki-67 is expressed in the nucleolus of interphase cells.
The Ki-67 gene contains 15 exons. The Ki-67 repeat region, within which there is a 22-amino acid Ki-67 motif, is encoded by exon 13. The shorter isoform lacks exon 7. Northern blot analysis revealed multiple transcripts ranging from approximately 8.9 to 12.5 kb in proliferating but not quiescent cells. Immunoblot analysis showed expression of 320- and 359-kD proteins. Sequence analysis predicted that the short-lived 2,896- and 3,256-amino acid protein isoforms contain potential nuclear targeting signals, over 200 potential phosphorylation sites, 19 N-myristoylation sites, 3 amidation sites, and numerous PEST sites. Antisense oligonucleotides inhibited cellular proliferation in a dose-dependent manner, suggesting that Ki-67 protein expression may be an absolute requirement for cell proliferation.
Product Form
A bivalent human recombinant Fab selected from the HuCAL® phage display library, expressed inE. coli.This Fab fragment is bivalent by dimerization of the bacterial alkaline phosphatase fusion protein. The antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This antibody is supplied lyophilised.
Storage
Prior to reconstitution store at +4oC.
After reconstitution store at -20oC.
Storage in frost-free freezers is not recommended. This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
12 months from date of reconstitution.
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
ELISA
Flow Cytometry
Functional Assays
Immunohistology - Frozen
Immunohistology - Paraffin(1)
1/50 - 1/200
Immunoprecipitation
Western Blotting
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
(1)
This product requires antigen retrieval using heat treatment prior to staining of paraffin sections.Sodium citrate buffer pH 6.0 is recommended for this purpose.
This product is suitable for use in direct ELISA applications. Due to the presence of bovine serum albumin (BSA), this antibody is unsuitable for use as a capture reagent in sandwich ELISA applications. This product is also available without BSA,please enquire.
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