PathScan? Phospho-HSP27 (Ser78) Sandwich ELISA Kit

• 产品编号：CST-7290C      品牌：CST       原厂货号：7290C
• 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's PathScan®Phospho-HSP27 (Ser78) Sandwich ELISA Kit可识别内源的磷酸化HSP27（Ser78）蛋白。使用Sandwich ELISA Kit #7290可以检测到紫外光处理后HeLa细胞中诱导产生的磷酸化HSP27（Ser78）。但Total HSP27 Sandwich ELISA Kit #7295检测出的HSP27总蛋白水平（磷酸化或非磷酸化）却没有变化（图1）。用紫外光处理后的COS细胞也可以得到类似的结果（数据未显示）。该试剂盒经过内部测试，可识别指定物种的蛋白质，也可能识别一些其它物种的同源性蛋白。CST's PathScan® Phospho-HSP27 (Ser78) Sandwich ELISA Kit是一个固相夹心酶联免疫吸附分析（ELISA）试剂盒，可以用来检测内源性的磷酸化HSP27（Ser78）蛋白。HSP27鼠单抗已经被包被于微孔中。在和细胞裂解液孵育时，HSP27蛋白（磷酸化和非磷酸化）将被包被抗体捕获。在充分冲洗后，Phospho-HSP27 (Ser78) Antibody被加入以捕获磷酸化HSP27（Ser78）蛋白。HRP-连接的抗鼠抗体#7074*将用来识别检测抗体。加入HRP底物TMB以显色。显色后的吸光度值高低正比于Ser78磷酸化HSP27的量的多少。* 试剂盒中的抗体仅对该试剂盒定制。热休克蛋白（HSP）27是小HSP之一，在多种细胞和组织中都有不同水平的组成性表达。像其它小HSP一样，HSP27的调节也发生在转录和翻译后（1）。应激反应中，HSP27表达增加了数倍以实现细胞对不利环境的抵抗。HSP27被MAPKAPK-2在Ser15、Ser78和Ser82磷酸化，作为p38 MAP激酶途径活化的结果（2,3）。HSP27磷酸化导致其四级结构改变，以至于从大分子量的同型多聚体变为二聚体和单体（4）。目前显示HSP27磷酸化和浓度增加可以调节肌动蛋白多聚化和再组织（5,6）。

1. Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373.
2. Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803.
3. Rouse, J. et al. (1994) Cell 78, 1027-1037.
4. Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956.
5. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
6. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Phospho-HSP27 (Ser78) Sandwich ELISA Kit detects endogenous levels of phospho-HSP27 (Ser78) protein. Using this Sandwich ELISA Kit #7290, a significant induction of phospho-HSP27 (Ser78) can be detected in HeLa cells treated with UV light. However, the level of total HSP27 (phospho and non-phospho), detected by the Total HSP27 Sandwich ELISA Kit #7295, remains unchanged (Figure 1). COS cells treated with UV light show similar results (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Description

CST's PathScan® Phospho-HSP27 (Ser78) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-HSP27 (Ser78) protein. HSP27 Mouse mAb has been coated onto the microwells. After incubation with cell lysates, HSP27 protein (phosphorylated and nonphosphorylated) is captured by the coated antibody. Following extensive washing, Phospho-HSP27 (Ser78) Antibody is added to detect the captured phospho- HSP27 (Ser78) protein. HRP-linked anti-rabbit antibody #7074* is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of phospho- HSP27 (Ser78) protein. 
 * Antibodies are custom formulations specific to the kit.

Background

Heat shock protein (HSP) 27 is one of the small HSPs that are constitutively expressed at different levels in various cell types and tissues. Like other small heat shock proteins, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the expression level of HSP27 increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78 and Ser82 by MAPKAP kinase 2 as a result of the activation of the p38 MAP kinase pathway (2,3). Phosphorylation of HSP27 causes a change in its tertiary structure, which shifts from large homotypic multimers to dimers and monomers (4). It has been shown that phosphorylation and increased concentration of HSP27 modulates actin polymerization and reorganization (5,6).

1. Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373.
2. Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803.
3. Rouse, J. et al. (1994) Cell 78, 1027-1037.
4. Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956.
5. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
6. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.

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).