Chk2, phosphorylated (Ser33/35) (Checkpoint Kinase 2, Rad 53)

Chk2 is the mammalian orthologue of the budding yeast Rad53 and fission yeast Cds1 checkpoint kinases. 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. After DNA damage by ionizing radiation (IR), UV irradiation or hydroxyurea treatment, Thr68 and other sites in this region become phosphorylated by ATM/ATR. 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 on residues Thr383 and Thr387 in the activation loop of the kinase domain.\n\nApplications: \nSuitable for use in Western Blot. Other applications not tested.\n\nRecommended Dilutions:\nWestern Blot: 1:1000\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

C4201-32

Size

100ul

Applications

WB

Hosts

Rabbit

Reactivities

Hum, NHP

Form

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.

P Type

Pab

Purity

Purifie by Protein A and peptide affinity chromatography.

Isotype

IgG

References

1. Allen, J.B. et al. (1994) Genes Dev. 8, 2401-2415. 2. Weinert, T.A. et al. (1994) Genes Dev. 8, 652-665. 3 .Murakami, H. and Okayama, H. (1995) Nature 374, 817-819. 4. Kastan, M.B. and Lim, D.S. (2000) Nat. Rev. Mol. Cell Biol. 1, 179-186. 5. Matsuoka, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 10389-10394. 6. Melchionna, R. et al. (2000) Nat. Cell Biol. 2, 762-765. 7. Ahn, J.Y. et al. (2000) Cancer Res. 60, 5934-5936. 8. Lee, C.H. and Chung, J.H. (2001) J. Biol. Chem. 276, 30537-30541.

Additional Info

Recognizes endogenous levels of human Chk2 only when phosphorylated at Ser33/35 at ~62kD. Does not crossreact with Chk2 phosphorylated at other sites. Species crossreactivity: monkey