PathScan? Total Chk2 Sandwich ELISA Antibody Pair

• 产品编号：CST-7090S      品牌：CST       原厂货号：7090S
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测抗体对
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描述：

抗体对的特异性和敏感性请参照对应的PathScan® Sandwich ELISA Kit。注意：本抗体对能够检测通过内部测试的特定种属蛋白，但也许能够检测其他种属的同源蛋白。相对于CST的PathScan® Total Chk2 Sandwich ELISA Kit #7045，PathScan® Total Chk2 Sandwich ELISA Antibody Pair提供了一个经济的选择。我们提供了捕获和检测抗体(100X储备液)和HRP-结合的二级抗体(1000X储备液)，针对4 x 96 孔ELISAs的试剂也非常充分。PathScan® Total Chk2 Sandwich ELISA Antibody Pair在含有PBS的96孔板中包被过夜。封闭后加入Chk2 detection antibody和anti-mouse IgG, HRP conjugated antibody，随后加入细胞裂解液。加入HRP底物(TMB)进行显色。显色过程中吸光度的强度和总Chk2蛋白的量成正比。 Chk2是哺乳动物中和芽殖酵母Rad53和裂殖酵母Cds1检验点激酶同源的蛋白(1-3)。Chk2的氨基末端结构域包含有一个七丝氨酸或者苏氨酸残基(Ser19, Thr26, Ser28, Ser33, Ser35, Ser50 and Thr68)组成的系列，每个系列紧跟着谷氨酸(SQ 或TQ模体)。这些地方是已知的ATM/ATR激酶磷酸化的首选作用位点(4,5)。电离辐射(IR)引起DNA损伤之后，UV照射处理或者羟基脲处理，这个区域的68位苏氨酸和其他位点被ATM/ATR磷酸化(5-7)。因此，SQ/TQ簇结构域似乎具有调节功能。第68位苏氨酸的磷酸化是后续激活步骤的先决条件，激活归因于激酶结构域激活回路中Chk2第383位和387位苏氨酸残基的自体磷酸化(8)。

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.

原厂资料：

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

Cell Signaling Technology's PathScan® Total Chk2 Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total Chk2 Sandwich ELISA Kit #7045. Capture and detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Chk2 capture antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a Chk2 detection antibody and an anti-mouse IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of total Chk2 protein.

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 on 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. 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.

注意事项：

Storage: Capture and detection antibodies are stored at 4ºC. 
Anti-Mouse IgG, HRP-linked Antibody is stored at -20ºC.