新的PathScan® 化学发光夹心ELISA试剂盒比传统的显色检测系统具有更宽的动态范围和更强的灵敏性.这些试剂盒提供小体积的微孔板，可以在使用小体积样本时保持强信号和灵敏性。

实验室的研发，验证和优化保证了ELISA试剂盒的高质量。

化学发光检测系统提供了最宽的动态范围和最好的检测灵敏度。

技术支持由开发和生产试剂盒的相同科学家提供。

12116      PathScan® Phospho-Acetyl-CoA Carboxylase (Ser79) Chemiluminescent Sandwich ELISA Kit

12321     PathScan® Phospho-Ret (panTyr) Chemiluminescent Sandwich ELISA Kit

7021       PathScan® Phospho-FLT3 (Tyr591) Chemiluminescent Sandwich ELISA Kit

7024       PathScan® Phospho-TrkA (Tyr490) Chemiluminescent Sandwich ELISA Kit

7027       PathScan® Phospho-c-Jun (Ser63) Chemiluminescent Sandwich ELISA Kit

7036       PathScan® Phospho-EGF Receptor (Tyr1068) Chemiluminescent Sandwich ELISA Kit

7084       PathScan® Total ALK Chemiluminescent Sandwich ELISA Kit

7087       PathScan® Phospho-TrkB (panTyr) Chemiluminescent Sandwich ELISA Kit

7132       PathScan® Total Akt1 Chemiluminescent Sandwich ELISA Kit

7134       PathScan® Phospho-Akt1 (Ser473) Chemiluminescent Sandwich ELISA Kit

7135       PathScan® Phospho-Akt (Thr308) Chemiluminescent Sandwich ELISA Kit

7149       PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit

7153       PathScan® Phospho-p70 S6 Kinase (Thr389) Chemiluminescent Sandwich ELISA Kit

7297       PathScan® Total EGF Receptor Chemiluminescent Sandwich ELISA Kit

7316       PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Chemiluminescent Sandwich ELISA Kit

7337       PathScan® Total S6 Ribosomal Protein Chemiluminescent Sandwich ELISA Kit

7792       PathScan® Phospho-AMPKα (Thr172) Chemiluminescent Sandwich ELISA Kit

7849       PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Chemiluminescent Sandwich ELISA Kit

7869       PathScan® Total SAPK/JNK Chemiluminescent Sandwich ELISA Kit

8026       PathScan® Phospho-Stat3 (Ser727) Chemiluminescent Sandwich ELISA Kit

Specificity / Sensitivity

PathScan® Total EGF Receptor Chemiluminescent Sandwich ELISA Kit #7297 detects endogenous levels of EGF receptor protein. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

PathScan® Total EGF Receptor Chemiluminescent Sandwich ELISA Kit #7297能够检测内源性水平的EGF受体蛋白。本试剂盒用来检测指示物种的蛋白，即通过内部测试确定的那些，也可以用来检测来自其他物种的同源蛋白。

Description

PathScan® Total EGF Receptor Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total EGF receptor protein. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using a smaller sample size. An EGF Receptor Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-EGF receptor proteins are captured by the coated antibody. Following extensive washing, EGF Receptor Rabbit mAb is added to detect both the captured phospho- and nonphospho-EGF receptor protein. Anti-rabbit IgG, HRP-linked Antibody #7074 is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of total EGF receptor protein. 
 Antibodies in kit are custom formulations specific to kit.

PathScan® Total EGF Receptor Chemiluminescent Sandwich ELISA Kit采用固相夹心酶联免疫吸附试验（ELISA），用化学发光法检测内源性水平的总表皮生长因子受体蛋白。化学发光酶联免疫吸附比传统的显色检测往往有更广阔的动态范围和更高的灵敏度。此化学发光ELISA提供在低量微孔板，提高信号灵敏度，同时使用一个较小的样本量。微孔包被EGF受体小鼠单克隆抗体。与细胞裂解物孵育后，磷酸化和非磷酸化-EGF受体蛋白质由包被的抗体捕获。充分洗涤后，加入EGF受体兔单克隆抗体，以检测捕获得磷酸和非磷酸-EGF受体蛋白。然后，使用抗兔IgG，HRP-标记的抗体＃7074，识别结合的检测抗体。加入化学发光试剂进行显色。发射光强度用相对光单位（RLU）测量，并与总EGF受体蛋白的量成正比。试剂盒中的抗体为试剂盒特别订制配方。

ELISA

Figure 1. The relationship between protein concentration of lysates from A-431 cells, untreated or treated with hEGF #8916, and immediate light generation with chemiluminescent substrate. After starvation, A-431 cells (85% confluence) were treated with hEGF (100 ng/ml, 5 min at 37°C) and then lysed. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.图1. A-431细胞裂解物的蛋白浓度之间的关系，细胞不处理或用hEGF #8916处理，立即产生光化学发光底物。饥饿处理后，A-431细胞（85%汇合）用hEGF处理 (100 ng/ml, 5 min at 37°C)，然后裂解。图中对应的阴影部分的面积图形显示了高灵敏度和低蛋白质浓度范围内的线性响应。

ELISA

Figure 2. The relationship between protein concentration of lysates prepared using H1975 cells, lysed with (phospho) and without (nonphospho) the addition of phosphatase inhibitors to the lysis buffer, and immediate light generation using chemiluminescent substrate.图2. H1975细胞裂解物蛋白浓度之间的关系，细胞用含（磷酸）和不含（非磷酸）添加磷酸酶抑制剂的裂解缓冲液进行裂解，立即产生光化学发光底物。

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).

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)。

7036 PathScan® Phospho-EGF Receptor (Tyr1068) Chemiluminescent Sandwich ELISA Kit

7074 Anti-rabbit IgG, HRP-linked Antibody

7187 PathScan® Phospho-EGF Receptor (Tyr1173) Sandwich ELISA Kit

7189 PathScan® Phospho-EGF Receptor (Tyr845) Sandwich ELISA Kit

7240 PathScan® Phospho-EGF Receptor (Tyr1068) Sandwich ELISA Kit

7250 PathScan® Total EGF Receptor Sandwich ELISA Kit

9803 Cell Lysis Buffer (10X)