PathScan? Phospho-EGF Receptor (panTyr) Sandwich ELISA Kit

• 产品编号：CST-7911C      品牌：CST       原厂货号：7911C
• 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：

PathScan^® Phospho-EGF Receptor (panTyr) Sandwich ELISA Kit能够检测内源性水平的人细胞酪氨酸磷酸化的EGF受体蛋白，见图1.试剂盒的灵敏性见图2.本试剂盒用来检测指定物种的蛋白，即通过内部测试确定的指定物种蛋白，也可能检测来自其他物种的同源蛋白。PathScan^® Phospho-EGF Receptor (panTyr) Sandwich ELISA Kit是固相夹心酶联免疫吸附试验（ELISA），能够检测内源性水平的酪氨酸磷酸化的EGF受体蛋白。EGF受体兔单克隆抗体包被微孔板。细胞裂解液孵育后，EGF受体蛋白（磷酸和非磷酸）由包被的抗体捕获。充分洗涤后，加入磷酸-酪氨酸兔检测单克隆抗体检测捕获的酪氨酸磷酸化的EGF受体蛋白。随后用抗鼠IgG, HRP-标记的抗体识别结合的检测蛋白。加入HRP底物TMB进行显色。显色的吸光度值与酪氨酸残基磷酸化的EGF受体蛋白的量成正比。试剂盒中的抗体是针对试剂盒定制的。表皮生长因子（EGF）受体是一种跨膜酪氨酸激酶，属于HER/ErbB蛋白家族。配体结合导致受体二聚化、自磷酸化、下游信号的激活、内化和溶酶体降解（1,2）。生长因子受体（EGFR）在激酶结构域酪氨酸845位点磷酸化，与稳定活化环、维持活性状态的酶和为底物蛋白提供一个结合表面有关（3,4）。c-Src参与表皮生长因子受体酪氨酸845位点的磷酸化(5)。PLCγ的SH2结构域结合磷酸化的酪氨酸992位点，导致PLCγ介导的下游信号的激活（6）。表皮生长因子受体酪氨酸1045位点磷酸化为接头蛋白c-Cbl创建一个主要的停靠点，导致受体EGFR激活后的泛素化和降解（7,8）。GRB2接头蛋白结合酪氨酸1068位点磷酸化的激活型表皮生长因子受体（9）。一对磷酸化表皮生长因子受体残基（Tyr1148和Tyr1173）为Shc支架蛋白提供了一个停靠点，两个位点都参与了MAP激酶信号激活过程（2）。表皮生长因子受体在特定的丝氨酸和苏氨酸残基的磷酸化，抑制表皮生长因子受体激酶的活性。表皮生长因子受体羧基末端残基丝氨酸（1046和1047位），都由钙调蛋白激酶II磷酸化；这些丝氨酸任何的突变，都导致EGFR酪氨酸自身磷酸化的上调（10）。

1. Hackel, P.O. et al. (1999) Curr Opin Cell Biol 11, 184-9.
2. Zwick, E. et al. (1999) Trends Pharmacol Sci 20, 408-12.
3. Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-4.
4. Hubbard, S.R. et al. (1994) Nature 372, 746-54.
5. Biscardi, J.S. et al. (1999) J Biol Chem 274, 8335-43.
6. Emlet, D.R. et al. (1997) J Biol Chem 272, 4079-86.
7. Levkowitz, G. et al. (1999) Mol Cell 4, 1029-40.
8. Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-66.
9. Rojas, M. et al. (1996) J Biol Chem 271, 27456-61.
10. Feinmesser, R.L. et al. (1999) J Biol Chem 274, 16168-73.

原厂资料：

Specificity / Sensitivity

PathScan^® Phospho-EGF Receptor (panTyr) Sandwich ELISA Kit detects endogenous levels of tyrosine-phosphorylated EGF receptor protein 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-EGF Receptor (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of tyrosine-phosphorylated EGF receptor protein. An EGF Receptor Rabbit mAb has been coated on the microwells. After incubation with cell lysates, EGF receptor protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-Tyrosine Mouse Detection mAb is added to detect captured tyrosine-phosphorylated EGF receptor 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 EGF receptor protein phosphorylated at tyrosine residues. 
 Antibodies in kit are custom formulations specific to kit.

Background

The epidermal growth factor (EGF) receptor is a transmembrane tyrosine kinase that belongs to the HER/ErbB protein family. Ligand binding results in receptor dimerization, autophosphorylation, activation of downstream signaling, internalization, and lysosomal degradation (1,2). Phosphorylation of EGF receptor (EGFR) at Tyr845 in the kinase domain is implicated in stabilizing the activation loop, maintaining the active state enzyme, and providing a binding surface for substrate proteins (3,4). c-Src is involved in phosphorylation of EGFR at Tyr845 (5). The SH2 domain of PLCγ binds at phospho-Tyr992, resulting in activation of PLCγ-mediated downstream signaling (6). Phosphorylation of EGFR at Tyr1045 creates a major docking site for the adaptor protein c-Cbl, leading to receptor ubiquitination and degradation following EGFR activation (7,8). The GRB2 adaptor protein binds activated EGFR at phospho-Tyr1068 (9). A pair of phosphorylated EGFR residues (Tyr1148 and Tyr1173) provide a docking site for the Shc scaffold protein, with both sites involved in MAP kinase signaling activation (2). Phosphorylation of EGFR at specific serine and threonine residues attenuates EGFR kinase activity. EGFR carboxy-terminal residues Ser1046 and Ser1047 are phosphorylated by CaM kinase II; mutation of either of these serines results in upregulated EGFR tyrosine autophosphorylation (10).

1. Hackel, P.O. et al. (1999) Curr Opin Cell Biol 11, 184-9.
2. Zwick, E. et al. (1999) Trends Pharmacol Sci 20, 408-12.
3. Cooper, J.A. and Howell, B. (1993) Cell 73, 1051-4.
4. Hubbard, S.R. et al. (1994) Nature 372, 746-54.
5. Biscardi, J.S. et al. (1999) J Biol Chem 274, 8335-43.
6. Emlet, D.R. et al. (1997) J Biol Chem 272, 4079-86.
7. Levkowitz, G. et al. (1999) Mol Cell 4, 1029-40.
8. Ettenberg, S.A. et al. (1999) Oncogene 18, 1855-66.
9. Rojas, M. et al. (1996) J Biol Chem 271, 27456-61.
10. Feinmesser, R.L. et al. (1999) J Biol Chem 274, 16168-73.

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).