Purified from mouse ascites fluids by affinity chromatography
Guaranteed Application *
WB, IHC, IP, FC
Suggested Dilutions
WB 1:1000~2000, IHC 1:150, FLOW 1:100, IP 2ug/500ul
Background
In response to DNA damage and replication blocks, cell cycle progression is halted through the control of critical cell cycle regulators. The protein encoded by this gene is a cell cycle checkpoint regulator and putative tumor suppressor. It contains a forkhead-associated protein interaction domain essential for activation in response to DNA damage and is rapidly phosphorylated in response to replication blocks and DNA damage. When activated, the encoded protein is known to inhibit CDC25C phosphatase, preventing entry into mitosis, and has been shown to stabilize the tumor suppressor protein p53, leading to cell cycle arrest in G1. In addition, this protein interacts with and phosphorylates BRCA1, allowing BRCA1 to restore survival after DNA damage. Mutations in this gene have been linked with Li-Fraumeni syndrome, a highly penetrant familial cancer phenotype usually associated with inherited mutations in TP53. Also, mutations in this gene are thought to confer a predisposition to sarcomas, breast cancer, and brain tumors. This nuclear protein is a member of the CDS1 subfamily of serine/threonine protein kinases. Three transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq].
Related Pathway
Cell cycle
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HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY CHEK2 (Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-CHEK2.
Western blot analysis of extracts (35ug) from 9 different cell lines by usin g anti-CHEK2 monoclonal antibody (HepG2: human; HeLa: human; SVT2: mouse; A549: human; COS7: monkey; Jurkat: human; MDCK: canine; PC12: rat; MCF7: human).
Immunoprecipitation(IP) of CHEK2 by using TrueMab monoclonal anti-CHEK2 antibodies (Negative control: IP without adding anti-CHEK2 antibody.). For each experiment, 500ul of DDK tagged CHEK2 overexpression lysates (at 1:5 dilution with HEK293T lysate), 2ug of anti-CHEK2 antibody and 20ul (0.1mg) of goat anti-mouse conjugated magnetic beads were mixed and incubated overnight. After extensive wash to remove any non-specific binding, the immuno-precipitated products were analyzed with rabbit anti-DDK polyclonal antibody.
HEK293T cells transfected with either RC201278 overexpress plasmid(Red) or empty vector control plasmid(Blue) were immunostained by anti-CHEK2 antibody(TA500384), and then analyzed by flow cytometry.
