PathScan? Phospho-Src (Tyr416) Sandwich ELISA Kit

• 产品编号：CST-7953C      品牌：CST       原厂货号：7953C
• 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-Src (Tyr416) Sandwich ELISA Kit能检测内源性Tyr416位点磷酸化的Src蛋白水平，如图1所示。该试剂盒不能与Src家族成员Yes, Lck, Lyn 和Hck发生交叉反应。其他家族成员还未经过测试。该试剂盒的敏感度如图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。但对其他物种中的同源蛋白可能也可以检测。PathScan® Phospho-Src (Tyr416) 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phospho-Src (Tyr416)的水平。先将phospho-Src rabbit antibody包被在微孔板上。与细胞裂解物孵育后，phospho-Src (Tyr416)便可被包被的抗体捕获。清洗彻底后，加入Src mouse detection antibody检测phospho-Src (Tyr416)的水平。再加入Anti-mouse， HRP偶联抗体作为二抗来识别之前绑定的检测抗体。最后加入HRP底物—— TMB，进行显色。产生的吸光值与phospho-Src (Tyr416)的数量成正比。Src家族蛋白酪氨酸激酶包括Src, Lyn, Fyn, Yes, Lck, Blk, Hck, 对调节真核细胞的生长和分化很重要(1)。Src的活性是通过两个酪氨酸位点的磷酸化来调控的，但是这两者的调控效应相反。激酶结构域的活化环中的Tyr416位点的磷酸化，能够上调酶活性；而蛋白羧基端尾部Tyr527位点被Csk磷酸化后，则会使酶的活性降低(2)。

1. Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609.
2. Hunter, T. (1987) Cell 49, 1-4.

原厂资料：

Specificity / Sensitivity

The PathScan® Phospho-Src (Tyr416) Sandwich ELISA Kit from Cell Signaling Technology detects endogenous levels of Src when phosphorylated at Tyr416, as shown in Figure 1. This kit does not cross-react with the Src family members Yes, Lck, Lyn or Hck. Other family members have not been tested. 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-Src (Tyr416) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Src (Tyr416). A phospho-Src rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-Src (Tyr416) is captured by the coated antibody. Following extensive washing, a Src mouse detection antibody is added to detect the captured phospho-Src (Tyr416). Anti-mouse, HRP-linked antibody is then used to recognize the bound detection antibody. The HRP substrate TMB is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-Src (Tyr416).

Background

The Src family of protein tyrosine kinases, which includes Src, Lyn, Fyn, Yes, Lck, Blk, and Hck, are important in the regulation of growth and differentiation of eukaryotic cells (1). Src activity is regulated by tyrosine phosphorylation at two sites, but with opposing effects. While phosphorylation at Tyr416 in the activation loop of the kinase domain upregulates enzyme activity, phosphorylation at Tyr527 in the carboxy-terminal tail by Csk renders the enzyme less active (2).

1. Thomas, S.M. and Brugge, J.S. (1997) Annu Rev Cell Dev Biol 13, 513-609.
2. Hunter, T. (1987) Cell 49, 1-4.

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).