Anti-Chk2 (Checkpoint Kinase 2, Rad 53), clone 2T8

Chk2 is the mammalian homologue 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) followed by glutamine (SQ or TQ motif). These are known to be preferred sites for phosphorylation by ATM/ATR kinases (4). Indeed, after DNA damage by ionizing radiation (IR), UV irradiation and DNA replication blocked by hydroxyurea, 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. Applications: Suitable for use in Western Blot, Immunoprecipitation and Immunocytochemistry. Other applications not tested. Recommended Dilutions: Western Blot: 0.1-1.0ug/ml detects CHK2 in lysates from Jurkat, HeLa, 3T3/A31, and A431 cells. Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-CHK2 (0.5ug/ml). Proteins were visualized using a IgG (HRP) goat anti-mouse secondary antibody and a chemiluminescence detection system. , Immunoprecipitation: Detects HK2 from HeLa cells. Reported by an independent lab., Immunocytochemistry: Detects CHK2 in HeLa cells. Reported by an independent lab., Optimal dilutions to be determined by the researcher. Positive Control Included: J9000: Jurkat cell lysate. Add 2.5ul of 2-mercaptoethanol/ 100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.