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Important Ordering Details

Custom Ordering Details: Product is assembled upon order to ensure maximum activity. United States: Please allow up to two weeks for your order to be processed and shipped. Outside of the United States: Please allow up to three weeks, depending on the country, for your order to be processed and shipped. When ordering five or more kits, please contact us for processing time and pricing at sales@cellsignal.com.

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REACTIVITY
Product Includes Volume Solution Color
Chk2 Ab Coated Microwells 96 assays
Chk2 Detection Ab 11 ml Green
Anti-mouse IgG, HRP-linked Antibody 11 ml Red
TMB Substrate 7004 11 ml Colorless
STOP Solution 7002 11 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) 9803 15 ml Yellowish

Product Description

The PathScan® Total Chk2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Chk2. A Chk2 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, Chk2 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Chk2 mouse detection antibody is added to the captured phospho and nonphospho Chk2 protein. Anti-mouse IgG, HRP-linked Antibody #7076 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total Chk2.

Antibodies in kit are custom formulations specific to kit.


Specificity / Sensitivity

CST's PathScan® Chk2 Sandwich ELISA Kit #7045 detects endogenous levels of total Chk2 protein (see Figure 1). The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.


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).


1.  Allen, J.B. et al. (1994) Genes Dev. 8, 2401-2415.

2.  Kastan, M.B. and Lim, D.S. (2000) Nat Rev Mol Cell Biol 1, 179-86.

3.  Weinert, T.A. et al. (1994) Genes Dev. 8, 652-665.

4.  Murakami, H. and Okayama, H. (1995) Nature 374, 817-819.

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.


Entrez-Gene Id 11200
Swiss-Prot Acc. O96017


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
PathScan® is a trademark of Cell Signaling Technology, Inc.