antibody

Chk2, phosphorylated, Thr68 (Checkpoint Kinase 2, Rad 53)

C4201-10

100 ul

polyclonal

domestic rabbit

nonconjugated

phosphorylated

Chk2, phosphorylated, Thr68 (Checkpoint Kinase 2, Rad 53)

rabbit

Pab

Phosphorylated

Antibodies

100 ul

IgG

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. The p53 tumor-suppressor gene integrates numerous signals that control cell life and death. Several novel molecules involved in p53 signaling, including CHK2, p53R2, p53AIP1, Noxa, PIDD, and PID/MTA2, were recently discovered. The checkpoint kinase CHK2 is the mammalian homologue of yeast Cds1/Rad53. In response to DNA damage, the checkpoint kinase ATM phosphorylates and activates CHK2, which in turn directly phosphorylates and activates p53. CHK2 serves as ATM downstream effector to mediate activation of p53. CHK2 also phosphorylates and activates BRCA1, the product of a tumor suppressor gene that is mutated in breast and ovarian cancer. Applications: Suitable for use in Western Blot, Immunocytochemistry, Immunofluorescence, Immunoprecipitation and Flow Cytometry. Other applications not tested. Recommended Dilutions: Western Blot: 1:1000 Immunofluorescence (IC): 1:100 Immunoprecipitation: 1:100 Flow Cytometry: 1:800 Optimal dilutions to be determined by the researcher. 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.

Hu Mk

Synthetic phosphopeptide corresponding to residues surrounding Thr68 of human Chk2.

Recognizes endogenous levels of human Chk2 only when phosphorylated at threonine 68 (62kD). Does not crossreact with Chk2 phosphorylated at other sites. Species Crossreactivity: monkey

Purified by Protein A and peptide affinity chromatography.

Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added.

Not Determined

Product Type: Pab Isotype: IgG Host: rabbit Source: human Concentration: Not Determined Form: Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol. No preservative added. Purity: Purified by Protein A and peptide affinity chromatography. Immunogen: Synthetic phosphopeptide corresponding to residues surrounding Thr68 of human Chk2. Specificity: Recognizes endogenous levels of human Chk2 only when phosphorylated at threonine 68 (62kD). Does not crossreact with Chk2 phosphorylated at other sites. Species Crossreactivity: monkey Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

FC IC IF IP WB

-20°C

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. Eastman, A. et al. (2002) Mol. Cancer Ther. 1, 1067–1078. 9. Kohn, E. A. et al. (2002) J. Biol. Chem. 277, 26553–26564.

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Flow cytometric analysis of HeLa cells, untreated (blue) and Uv-treated (green), using C4201-10.