Phospho-EGF Receptor Pathway Antibody Sampler Kit

• 产品编号：CST-9789S      品牌：Cell Signaling Technology       原厂货号：9789S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

磷酸化EEGFR通路小包装试剂盒中的每一个抗体都能够识别其特定位点的磷酸化位点。磷酸化EGFR（Tyr1068) (D7A5) XP®兔单抗可能与其他的酪氨酸磷酸化蛋白有微弱的交叉反应。磷酸化Gab1 (Tyr627) (C32H2)兔单抗可能与磷酸化的Gab2, Gab3或活化的受体酪氨酸激酶产生交叉反应。磷酸化Shc (Tyr239/240)抗体可能与活化的EGF受体蛋白产生交叉反应。磷酸化多克隆抗体是通过用合成磷酸化多肽免疫动物而生成，该磷酸化多肽根据人c-Cbl 的Tyr774位点，人Shc的Tyr239/240位点，以及人PLCγ1的Tyr783位点周围的残基序列设计。多抗的纯化采用蛋白A和多肽亲和层析法进行。兔单克隆抗体是通过用合成的磷酸化多肽免疫动物而生成，该磷酸化多肽根据人EGFR的Tyr1068，Stat5a的Tyr694，人Gab1的Tyr627，人Akt的Ser473，以及人p44 MAP激酶的Thr202/Tyr204位点周围的残基序列设计。磷酸化EGFR通路小包装试剂盒为检测EGFR通路中的多个成员的激活状态提供了一种经济的方式，包括检测EGFR, Stat5, c-Cbl, Shc, Gab1, PLCγ1, Akt和p44/42 MAPK的磷酸化水平。该试剂盒包括足够进行4次WB试验的一抗和二抗。表皮生长因子（EGF）受体是一个属于HER/ErbB蛋白家族的跨膜酪氨酸激酶。配体的结合导致了受体的二聚化，自磷酸化，下游信号分子的激活，内化以及溶酶体降解 (1,2)。表皮生长因子受体（EGFR）在激酶域Tyr845位点的磷酸化与稳定激活回路有关，并且维持了酶的激活状态，为底物蛋白提供了可结合的表位(3,4)。c-Src与 EGFR 在 Tyr845位点的磷酸化有关(5)。PLCγ的SH2结构域结合到磷酸化的Tyr992位点，将引起PLCγ介导的下游信号通路的激活(6)。EGFR Tyr1045位点的磷酸化则为c-Cbl生成一个主要的停靠位点，这个衔接蛋白将导致受体的泛素化以及EGFR活化后的降解(7,8)。GRB2衔接蛋白则结合到活化后的EGFR磷酸化的Tyr1068位点(9)。一对磷酸化EGFR残基(Tyr1148 and Tyr1173)为Shc支架蛋白提供了停靠点，这两个位点都与MAPK激酶通路的激活有关(2)。EGFR在特定的丝氨酸和苏氨酸残基上的磷酸化减弱了EGFR激酶的活性。EGFR羧基末端残基Ser1046 和 Ser1047被CaM 激酶II磷酸化，这两个丝氨酸任何一个突变都将导致EGFR酪氨酸自身磷酸化的上调(10)。

原厂资料：

Specificity / Sensitivity

Each antibody in the Phospho-EGF Receptor Pathway Sampler Kit recognizes the phosphorylated form of its specific target. Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb may cross-react weakly with other tyrosine-phosphorylated proteins. Phospho-Gab1 (Tyr627) (C32H2) Rabbit mAb may cross-react with phosphorylated Gab2, Gab3, or activated receptor tyrosine kinases. Phospho-Shc (Tyr239/240) Antibody may cross-react with activated EGF receptor protein.

Source / Purification

Activation state polyclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr774 of human c-Cbl, Tyr239/240 of human Shc and Tyr783 of human PLCγ1. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Rabbit monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Tyr1068 of human EGF receptor, Tyr694 of Stat5a, Tyr627 of human Gab1, Ser473 of human Akt and Thr202/Tyr204 of human p44 MAP kinase.

Description

The Phospho-EGF Receptor Pathway Sampler Kit provides an economical means to evaluate the activation status of multiple members of the EGF receptor pathway, including phosphorylated EGF receptor, Stat5, c-Cbl, Shc, Gab1, PLCγ1, Akt and p44/42 MAPK. The kit includes enough primary and secondary antibodies to perform four western blot experiments. 

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 c-Cbl, an adaptor protein that leads 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) provides 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).

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibody