PathScan? Phospho-Syk (Tyr525/526) Sandwich ELISA Kit

• 产品编号：CST-7970C      品牌：CST       原厂货号：7970C
• 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：

CST的PathScan® Phospho-Syk (Tyr525/526) Sandwich ELISA Kit #7970检测内源水平在Tyr525/526位点磷酸化的Syk蛋白。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-Syk (Tyr525/526) 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tyr525/526位点磷酸化的Syk蛋白。phospho-Syk (Tyr525/526)鼠抗包裹到微孔的外表面。然后与细胞裂解液孵育，在tyr525/526位点磷酸化的Syk蛋白能够被抗体捕获。经过严格的冲洗后，Syk鼠单检测抗体加入用来检测捕捉到的Syk蛋白。HRP连接抗鼠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Tyr525/526位点磷酸化的Syk蛋白的量。*试剂盒中的抗体是经过客户证实试剂盒特异的。Syk作为酪氨酸蛋白激酶在造血细胞的胞内信号转导中发挥重要作用(1-3)。 Syk能够与免疫受体的酪氨酸介导的活性区域(ITAMs)中的胞质部分结合(4)。它能偶联激活了的免疫受体进行下游的信号转导，而引起一系列的细胞应答，如细胞增殖、分化以及吞噬(4)。有证据显示Syk在无免疫反应的细胞中也有作用，Syk在人类的乳腺癌中是个潜在的肿瘤抑制因子(5)。 Syk蛋白的SH2-激酶结合区域中Tyr323位点是个磷酸化负调控位点。磷酸化的Tyr323为 Cbl的TKB区域提供了结合位点(6,7)。Syk的Tyr352位点参与了与 PLC-γ1 的结合 (8)。 Tyr525位点和526位点位于Syk的激酶区域的活性环上。磷酸化人源的Syk的Tyr525/526位点(等同于小鼠 Syk的 Tyr519/520位点) 对 Syk 的功能有重要的影响(9)。

1. Cheng, A.M. and Chan, A.C. (1997) Curr Opin Immunol 9, 528-33.
2. Kurosaki, T. (1997) Curr Opin Immunol 9, 309-18.
3. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
4. Turner, M. et al. (2000) Immunol Today 21, 148-54.
5. Coopman, P.J. et al. (2000) Nature 406, 742-7.
6. Deckert, M. et al. (1998) J Biol Chem 273, 8867-74.
7. Rao, N. et al. (2001) EMBO J 20, 7085-95.
8. Law, C.L. et al. (1996) Mol Cell Biol 16, 1305-15.
9. Zhang, J. et al. (2000) J Biol Chem 275, 35442-7.

原厂资料：

Specificity / Sensitivity

CST's PathScan® Phospho-Syk (Tyr525/526) Sandwich ELISA Kit #7970 detects endogenous levels of Syk protein when phosphorylated at Tyr525/526. 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-Syk (Tyr525/526) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Syk when phosphorylated at tyrosine 525/526. A phospho-Syk (Tyr525/526) Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Syk phosphorylated at Tyrosine525/526 is captured by the coated antibody. Following extensive washing, a Syk mouse monoclonal detection antibody is added to detect the captured Syk protein. HRP-linked Anti-mouse Ab 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 Syk phosphorylated on Tyr525/526. 
 Antibodies in kit are custom formulations specific to kit.

Background

Syk is a protein tyrosine kinase that plays an important role in intracellular signal transduction in hematopoietic cells (1-3). Syk interacts with immunoreceptor tyrosine-based activation motifs (ITAMs) located in the cytoplasmic domains of immune receptors (4). It couples the activated immunoreceptors to downstream signaling events that mediate diverse cellular responses, including proliferation, differentiation and phagocytosis (4). There is also evidence of a role for Syk in nonimmune cells, and Syk is a potential tumor suppressor in human breast carcinomas (5). Tyr323 is a negative regulatory phosphorylation site within the SH2-kinase linker region in Syk. Phosphorylation of Tyr323 provides a direct binding site to the TKB domain of Cbl (6,7). Tyrosine 352 of Syk is involved in the association of PLC-γ1 (8). Tyrosines 525 and 526 are located in the activation loop of the Syk kinase domain, and phosphorylation of Tyr525/526 of human Syk (equivalent to the Tyr519/520 of mouse Syk) is essential for Syk function (9).

1. Cheng, A.M. and Chan, A.C. (1997) Curr Opin Immunol 9, 528-33.
2. Kurosaki, T. (1997) Curr Opin Immunol 9, 309-18.
3. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
4. Turner, M. et al. (2000) Immunol Today 21, 148-54.
5. Coopman, P.J. et al. (2000) Nature 406, 742-7.
6. Deckert, M. et al. (1998) J Biol Chem 273, 8867-74.
7. Rao, N. et al. (2001) EMBO J 20, 7085-95.
8. Law, C.L. et al. (1996) Mol Cell Biol 16, 1305-15.
9. Zhang, J. et al. (2000) J Biol Chem 275, 35442-7.

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).