Description
Product Usage Information
Western Blotting: 1:1000
Storage
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at -20°C. Do not aliquot the antibody.
Protocol
Available protocols: Western Blotting
Specificity / Sensitivity
Phospho-Chk2 (Ser516) Antibody detects endogenous levels of Chk2 only when phosphorylated at serine 516. This antibody does not cross-react with Chk2 phosphorylated at other sites.
Species Reactivity: Human
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser516 of human Chk2. Antibodies are purified by protein A and peptide affinity chromatography.
Background
Chk2 is the mammalian orthologue of the budding yeast Rad53 and fission yeast Cds1 checkpoint kinases (1-3). The amino-terminal domain of Chk2 contains a series of seven serine or threonine residues (Ser19, Thr26, Ser28, Ser33, Ser35, Ser50, and Thr68) each followed by glutamine (SQ or TQ motif). These are known to be preferred sites for phosphorylation by ATM/ATR kinases (4,5). After DNA damage by ionizing radiation (IR), UV irradiation, or hydroxyurea treatment, Thr68 and other sites in this region become phosphorylated by ATM/ATR (5-7). The SQ/TQ cluster domain, therefore, seems to have a regulatory function. Phosphorylation at Thr68 is a prerequisite for the subsequent activation step, which is attributable to autophosphorylation of Chk2 at residues Thr383 and Thr387 in the activation loop of the kinase domain (8). Chk2 autophosphorylation at Ser516 is important for optimal Chk2 function, and a Ser516Ala mutant Chk2 is defective in IR-induced apoptosis (9).
Alternate Names
Cds1; Cds1 homolog; Checkpoint kinase 2; checkpoint-like protein CHK2; CHEK2; CHK2; CHK2 checkpoint homolog; CHK2 checkpoint homolog (S. pombe); hCds1; HuCds1; LFS2; PP1425; protein kinase CHK2; RAD53; Serine/threonine-protein kinase Chk2
