PathScan? Phospho-Stat1 (Tyr701) Sandwich ELISA Kit

• 产品编号：CST-7234C      品牌：CST       原厂货号：7234C
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测试剂盒
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描述：

CST的 PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit检测内源水平在Tyr701位点磷酸化的Stat1蛋白，如图1所示。在HeLa细胞中，用 Phospho-Stat1 (Tyr701) Sandwich ELISA Kit #7234检测经IFN-γ处理诱导Tyr701位点磷酸化Stat1蛋白。通过Western blot表明总Stat1（磷酸化和非磷酸化）蛋白并未受到影响。此试剂盒能够检测已经经内部严格测试的物种的蛋白，但是也不排除检测到其它物种中同源性的蛋白。PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit是固相夹心酶联免疫吸收剂检测试剂盒(ELISA)，它能够检测在Tyr701位点磷酸化的Stat1蛋白。Stat1兔抗包裹到微孔的外表面。然后与细胞裂解液孵育，Stat1(磷酸化和非磷酸化)蛋白都能够被抗体捕获。经过严格的冲洗后，生物素化的phospho-Stat1(Tyr701)兔检测抗体加入用来检测捕捉到的Tyr701位点磷酸化的Stat1蛋白。HRP连接链霉亲和素用于识别检测抗体的结合。HRP的底物TMB加入进行颜色反应。对反应的颜色，吸光度的数值按比例换算成在Tyr701位点磷酸化Stat1蛋白的量。*试剂盒中的抗体是经过客户验证的特异性抗体。Stat1转录因子可以被一系列配体所激活(1)， 其中对IFN-α和 IFN-γ的应答非常重要(2,3)。 在 Tyr701位点的磷酸化诱导 Stat1形成二聚体, 入核后与DNA结合(4)。 Stat1 蛋白是以Stat1α (91 kDa) 和 剪接变体Stat1β (84 kDa)二聚体的形式存在。在很多细胞中，这两种形式都是通过IFN-α激活。但是只有 Stat1α是被 IFN-γ激活。Stat1这种不正常的激活方式存在于很多肿瘤(5)。除了酪氨酸磷酸化，Stat1 也能通过p38有丝分裂激活蛋白激酶依赖(MAPK)依赖的通路在Ser727位点发生磷酸化而被激活，从而对IFN-α和其它的细胞应激产生应答(6)。丝氨酸磷酸化对最大化的诱导Stat1介导的基因激活也是必需的。

1. Heim, M.H. (1999) J Recept Signal Transduct Res 19, 75-120.
2. Durbin, J.E. et al. (1996) Cell 84, 443-50.
3. Meraz, M.A. et al. (1996) Cell 84, 431-42.
4. Ihle, J.N. et al. (1994) Trends Biochem Sci 19, 222-7.
5. Frank, D.A. (1999) Mol Med 5, 432-56.
6. Wen, Z. et al. (1995) Cell 82, 241-50.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit detects endogenous levels of Phospho-Stat1 when phosphorylated at Tyr701. As shown in Figure 1, a significant induction of Stat1 phosphorylation at Tyr701 can be detected in HeLa cells following treatment with Interferon-γ (IFN-γ) using the Phospho-Stat1 (Tyr701) Sandwich ELISA Kit #7234. The level of total Stat1 (phospho and nonphospho) remains unchanged as shown by Western analysis. 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-Stat1 (Tyr701) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Stat1 when phosphorylated at Tyr701. A Stat1 Rabbit Antibody has been coated onto the microwells. After incubation with cell lysates, Stat1 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, biotinylated Phospho-Stat1 (Tyr701) Rabbit Detection Antibody is added to detect phosphorylation of Tyr701 on the captured Stat1 protein. HRP-linked streptavidin 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 Stat1 phosphorylated at Tyr701. 
 Antibodies in kit are custom formulations specific to kit.

Background

The Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation, and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation.

1. Heim, M.H. (1999) J Recept Signal Transduct Res 19, 75-120.
2. Durbin, J.E. et al. (1996) Cell 84, 443-50.
3. Meraz, M.A. et al. (1996) Cell 84, 431-42.
4. Ihle, J.N. et al. (1994) Trends Biochem Sci 19, 222-7.
5. Frank, D.A. (1999) Mol Med 5, 432-56.
6. Wen, Z. et al. (1995) Cell 82, 241-50.

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).