PathScan? Phospho-Lck (Tyr505) Sandwich ELISA Kit

• 产品编号：CST-7941C      品牌：CST       原厂货号：7941C
• 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-Lck (Tyr505) Sandwich ELISA Kit检测内源在Tyr505位点磷酸化的Lck，如图1所示。此试剂盒不与Src家族的其他成员Yes, Src, Lyn 或 Hck交叉反应。其他家族的成员尚未测试。试剂盒的灵敏性如图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-Lck (Tyr505) Sandwich ELISA Kit是固相夹心酶联免疫吸收剂检测试剂盒(ELISA)，它能够检测内源在Tyr505位点磷酸化的Lck蛋白。phospho-Lck 兔抗包裹到微孔的外表面。然后与细胞裂解液孵育，phospho-Lck (Tyr505)蛋白能够被抗体捕获。经过严格的冲洗后, Lck鼠检测抗体加入用来检测捕捉到的phospho-Lck (Tyr505)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加入进行显色反应。对反应的颜色，吸光度的数值按比例换算成phospho-Lck (Tyr505)蛋白的量。Lck 属于Src样无受体酪氨酸激酶家族，具有典型的Src家族激酶的结构: 一个独特的N-末端域(Src同源4结构域, SH4) 然后是一个SH3 结构域, 一个SH2 结构域, 激酶结构域(SH1)和C-末端负调节结构域(1)。Lck的活性受到SH2和SH3区域的相互作用和在活性环内酪氨酸磷酸化的状态的共同调控(2,3)。受到刺激后，Lck蛋白被招募到T细胞受体(TCR)复合体上并激活下游酪氨酸激酶起始的T细胞信号通路(4)。Lck被发现也参与到线粒体凋亡信号通路的调控中，可能也对一些抗癌药物诱导凋亡作出反应(5,6)。

1. Palacios, E.H. and Weiss, A. (2004) Oncogene 23, 7990-8000.
2. Mustelin, T. and Taskén, K. (2003) Biochem J 371, 15-27.
3. Gervais, F.G. et al. (1993) Mol Cell Biol 13, 7112-21.
4. Straus, D.B. and Weiss, A. (1992) Cell 70, 585-93.
5. Belka, C. et al. (2003) Oncogene 22, 176-85.
6. Gruber, C. et al. (2004) Biochem Pharmacol 67, 1859-72.

原厂资料：

Specificity / Sensitivity

The PathScan® Phospho-Lck (Tyr505) Sandwich ELISA Kit from Cell Signaling Technology detects endogenous levels of Lck when phosphorylated at Tyr505, as shown in Figure 1. This kit does not cross-react with the Src family members Yes, Src, 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-Lck (Tyr505) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Lck (Tyr505). A phospho-Lck rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-Lck (Tyr505) is captured by the coated antibody. Following extensive washing, a Lck mouse detection mAb is added to detect the captured phospho-Lck (Tyr505). 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-Lck (Tyr505).

Background

Lck belongs to the Src-like non-receptor tyrosine kinase family with the typical Src family kinase structure: a unique amino terminal domain (Src homology 4 domain, SH4) followed by an SH3 domain, an SH2 domain, a kinase domain (SH1), and a carboxy-terminal negative regulatory domain (1). Lck activity is controlled by the interactions of SH2 and SH3 domains as well as tyrosine phosphorylation status of the activation loop (2,3). Lck is recruited to the T cell receptor (TCR) complex upon stimulation and activates downstream tyrosine kinases to initiate T cell signaling (4). Lck is also found to be involved in the regulation of mitochondrial apoptosis pathways and may be responsible for some anticancer drug induced apoptosis (5,6).

1. Palacios, E.H. and Weiss, A. (2004) Oncogene 23, 7990-8000.
2. Mustelin, T. and Taskén, K. (2003) Biochem J 371, 15-27.
3. Gervais, F.G. et al. (1993) Mol Cell Biol 13, 7112-21.
4. Straus, D.B. and Weiss, A. (1992) Cell 70, 585-93.
5. Belka, C. et al. (2003) Oncogene 22, 176-85.
6. Gruber, C. et al. (2004) Biochem Pharmacol 67, 1859-72.

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).