PathScan? Phospho-Stat3 (Ser727) Sandwich ELISA Kit

• 产品编号：CST-7995C      品牌：CST       原厂货号：7995C
• 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-Stat3 (Ser727) Sandwich ELISA Kit检测内源水平在Ser727位点磷酸化人Stat3蛋白，如图1所示。试剂盒的灵敏度如图2所示。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-Stat3 (Ser727) 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Ser727位点磷酸化的Stat3蛋白。Stat3鼠抗包裹到微孔的外表面。然后与细胞裂解液孵育，Stat3(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，phospho-Stat3 (Ser727)鼠检测抗体加入用来检测捕捉到的phospho-Stat3蛋白。抗鼠IgG，HRP连接抗体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，光密度的数值按比例换算成在Ser727位点磷酸化Stat3蛋白的量。*试剂盒中的抗体是经过客户证实试剂盒特异的。Stat3 转录因子是细胞因子和生长因子受体信号通路中重要的信号分子(1)，并且对于小鼠胎儿发育是必须的(2)。Stat3 在很多的人类肿瘤中是组成性激活的(3,4)并有潜在的支配原发性肿瘤的可能(5) ，而且也具有抗凋亡活性(3)。Stat3通过Tyr705位点的磷酸化而激活，从而导致它的二聚化，入核后与DNA结合(6,7)。转录活性似乎受到 MAPK 或 mTOR 信号通路介导的Stat3 Ser727位点的磷酸化的调控(8,9)。Stat3不同剪切形式的表达反应了其生物学功能，Stat3α (86 kDa) 和 Stat3β (79 kDa) 形式的表达比例依赖于细胞类型、配体方向或者细胞发育的成熟阶段(10)。值得注意的是，Stat3β蛋白在C-末端转录活性区域缺少丝氨酸磷酸化位点(8)。

1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
7. Ihle, J.N. (1995) Nature 377, 591-4.
8. Wen, Z. et al. (1995) Cell 82, 241-50.
9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

原厂资料：

Specificity / Sensitivity

PathScan® Phospho-Stat3 (Ser727) Sandwich ELISA Kit detects endogenous levels of Stat3 protein phosphorylated at Ser727 in human cells, as shown in Figure 1. The 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-Stat3 (Ser727) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Stat3 protein phosphorylated at Ser727. A Stat3 rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Stat3 (phospho or nonphospho) protein is captured by the coated antibody. Following extensive washing, a phospho-Stat3 (Ser727) mouse mAb is added to detect the captured phospho-Stat3 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 this developed color is proportional to the quantity of Stat3 phosphorylated at Ser727. 
 Antibodies in kit are custom formulations specific to kit.

Background

The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).

1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
7. Ihle, J.N. (1995) Nature 377, 591-4.
8. Wen, Z. et al. (1995) Cell 82, 241-50.
9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

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).