PathScan? Phospho-eIF2α (Ser51) Sandwich ELISA Kit

• 产品编号：CST-7286C      品牌：CST       原厂货号：7286C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
• 应用分类：

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描述：

PathScan® Phospho-eIF2α (Ser51) Sandwich ELISA Kit #7286能够检测内源性Ser51位点磷酸化的eIF2α水平，如图1所示。试剂盒敏感性如图2所示。经内部测试确认，本试剂盒可以检测到所列物种中的蛋白。但也可能检测出其他物种中的同源蛋白。PathScan® Phospho-eIF2α (Ser51) Sandwich ELISA Kit是通过固相夹心酶联免疫吸附法（ELISA）来检测内源性的内源性Ser51位点磷酸化的eIF2α水平的。先将兔源eIF2α抗体包被在微孔板上。与细胞裂解物孵育后，eIF2α便可被包被的抗体捕获。清洗彻底后，加入phospho-eIF2α (Ser51)鼠源检测抗体来检测phospho-eIF2α蛋白。再加入HRP标记抗小鼠 IgG作为二抗来识别之前结合的检测抗体。最后加入HRP底物TMB进行显色。产生的吸光值与Ser51位点磷酸化的eIF2α的数量成正比。试剂盒中抗体是专为本试剂盒特制的。研究证明在不同的应激条件下真核生物起始因子2 (eIF2) α亚型的磷酸化可下调蛋白质的合成。eIF2蛋白与GTP以及起始tRNA（Met-tRNAi）结合，然后将Met-tRNA转移到40S核糖体亚基上共同形成43S前起始复合物(1,2)。eIF2通过eIF2B催化反应将GTP交换GDP从而促进新一轮翻译的起始(1,2)。病毒感染(PKR)、内质网应激（endoplasmic reticulum stress）(PERK/PEK)、氨基酸饥饿(GCN2)或血红素缺乏症(HRI)能够激活相关的激酶，这些激酶可以使eIF2的α亚基磷酸化(3,4)。这个磷酸化稳固eIF2-GDP-eIF2B复合物，同时期抑制eIF2B的降解。通过IFN-γ 和TNF-α的诱导的PKR能有效的诱导eIF2α蛋白在Ser51位点磷酸化(5,6)。

1. Kimball, S.R. (1999) Int. J. Biochem. Cell Biol. 31, 25-29.
2. De Haro, C. et al. (1996) FASEB J. 10, 1378-1387.
3. Kaufman, R.J. (1999) Genes Dev. 13, 1211-1233.
4. Sheikh, M.S. and Fornace Jr., A.J. (1999) Oncogene 18, 6121-6128.
5. Cheshire, J.L. et al. (1999) J. Biol. Chem. 274, 4801-4806.
6. Zamanian-Daryoush, M. et al. (2000) Mol. Cell. Biol. 20, 1278-1290.

原厂资料：

Specificity / Sensitivity

The PathScan® Phospho-eIF2α (Ser51) Sandwich ELISA Kit #7286 detects endogenous levels of eIF2α protein when phosphorylated at Ser51 as shown in Figure 1. 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.

Description

The PathScan® Phospho-eIF2α (Ser51) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of eIF2α phosphorylated at Ser51. A eIF2α rabbit antibody has been coated onto the microwells. After incubation with cell lysates, eIF2α protein is captured by the coated antibody. Following extensive washing, a phospho-eIF2α (Ser51) mouse detection antibody is added to detect captured phospho-eIF2α protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound mouse 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 eIF2α phosphorylated at Ser51. 
 Antibodies in kit are custom formulations specific to kit.

Background

Phosphorylation of the eukaryotic initiation factor 2 (eIF2) α subunit is a well-documented mechanism to downregulate protein synthesis under a variety of stress conditions. eIF2 binds GTP and Met-tRNAi and transfers Met-tRNA to the 40S subunit to form the 43S preinitiation complex (1,2). eIF2 promotes a new round of translation initiation by exchanging GDP for GTP, a reaction catalyzed by eIF2B (1,2). Kinases that are activated by viral infection (PKR), endoplasmic reticulum stress (PERK/PEK), amino acid deprivation (GCN2), or heme deficiency (HRI) can phosphorylate the α subunit of eIF2 (3,4). This phosphorylation stabilizes the eIF2-GDP-eIF2B complex and inhibits the turnover of eIF2B. Induction of PKR by IFN-γ and TNF-α induces potent phosphorylation of eIF2α at Ser51 (5,6).

1. Kimball, S.R. (1999) Int. J. Biochem. Cell Biol. 31, 25-29.
2. De Haro, C. et al. (1996) FASEB J. 10, 1378-1387.
3. Kaufman, R.J. (1999) Genes Dev. 13, 1211-1233.
4. Sheikh, M.S. and Fornace Jr., A.J. (1999) Oncogene 18, 6121-6128.
5. Cheshire, J.L. et al. (1999) J. Biol. Chem. 274, 4801-4806.
6. Zamanian-Daryoush, M. et al. (2000) Mol. Cell. Biol. 20, 1278-1290.

注意事项：

* 12 8-well modules -Each module is designed to break apart for 8 tests.
**Kit should be stored at 4°C with the exception of 10X Cell Lysis Buffer, which is stored at –20°C (packaged separately).