The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
Use a concentration of 1 µg/ml.
FunctionRegulates cell cycle checkpoints and apoptosis in response to DNA damage, particularly to DNA double-strand breaks. Inhibits CDC25C phosphatase by phosphorylation on 'Ser-216', preventing the entry into mitosis. May also play a role in meiosis. Regulates the TP53 tumor suppressor through phosphorylation at 'Thr-18' and 'Ser-20'.
Tissue specificityHigh expression is found in testis, spleen, colon and peripheral blood leukocytes. Low expression is found in other tissues.
Involvement in diseaseDefects in CHEK2 are associated with Li-Fraumeni syndrome 2 (LFS2) [MIM:609265]; a highly penetrant familial cancer phenotype usually associated with inherited mutations in p53/TP53. Defects in CHEK2 may be a cause of susceptibility to prostate cancer (PC) [MIM:176807]. It is a malignancy originating in tissues of the prostate. Most prostate cancers are adenocarcinomas that develop in the acini of the prostatic ducts. Other rare histopathologic types of prostate cancer that occur in approximately 5% of patients include small cell carcinoma, mucinous carcinoma, prostatic ductal carcinoma, transitional cell carcinoma, squamous cell carcinoma, basal cell carcinoma, adenoid cystic carcinoma (basaloid), signet-ring cell carcinoma and neuroendocrine carcinoma. Defects in CHEK2 are found in some patients with osteogenic sarcoma (OSRC) [MIM:259500].
Sequence similaritiesBelongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CHK2 subfamily. Contains 1 FHA domain. Contains 1 protein kinase domain.
Post-translational modificationsPhosphorylated by PLK4.
Cellular localizationNucleus; Nucleus. Isoform 10 is present throughout the cell and Nucleus > PML body. Nucleus > nucleoplasm. Recruited into PML bodies together with TP53.
Western blot - Chk2 (phospho S28) antibody (ab26340)
All lanes : Anti-Chk2 (phospho S28) antibody (ab26340) at 1 µg/ml
Lane 1 : HeLa Gamma Irradiated Whole Cell Lysate Lane 2 : HeLa Gamma Irradiated Whole Cell Lysate with Human Chk2 (phospho S28) peptide (ab28701) at 1 µg/ml Lane 3 : HeLa Gamma Irradiated Whole Cell Lysate with Human Chk2 peptide (ab28702) at 1 µg/ml
ICC/IF image of ab26340 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab26340, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Immunocytochemistry/ Immunofluorescence - Chk2 (phospho S28) antibody (ab26340)This image is courtesy of an Abreview submitted by Dr Kirk McManus
ab26340 (1/500) staining Chk2 (phospho S28) in HeLa cells. Cells were pre-treated with Bleomycin for 2 hours, paraformaldehyde fixed and permeabilised with 0.5% Triton X100/PBS. Colocalisation with sites of double-strand DNA repair was confirmed through the use of an anti-gamma H2AX antibody (ab18311). Please refer to abreview for further experimental details.