| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Phospho-Chk2 (Thr68) Rabbit mAb Coated Microwells | 78149 | 96 tests |
|
+4C |
| Chk2 Mouse Detection mAb | 4831 | 1 ea |
|
+4C |
| Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) | 13304 | 1 ea |
|
+4C |
| Detection Antibody Diluent | 13339 | 11 ml |
|
+4C |
| HRP Diluent | 13515 | 11 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| ELISA Sample Diluent | 11083 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The PathScan® Phospho-Chk2 (Thr68) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Chk2 when phosphorylated at Thr68. A phospho-Chk2 (Thr68) rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-Chk2 protein is captured by the coated antibody. Following extensive washing, a Chk2 mouse detection antibody is added to detect the captured Chk2 protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of Chk2 phosphorylated at Thr68.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
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).
- Allen, J.B. et al. (1994) Genes Dev. 8, 2401-2415.
- Weinert, T.A. et al. (1994) Genes Dev. 8, 652-665.
- Murakami, H. and Okayama, H. (1995) Nature 374, 817-819.
- Kastan, M.B. and Lim, D.S. (2000) Nat. Rev. Mol. Cell Biol. 1, 179-186.
- Matsuoka, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 10389-10394.
- Melchionna, R. et al. (2000) Nat. Cell Biol. 2, 762-765.
- Ahn, J.Y. et al. (2000) Cancer Res. 60, 5934-5936.
- Lee, C.H. and Chung, J.H. (2001) J. Biol. Chem. 276, 30537-30541.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
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