| 货号 | 54359S |
| 反应种属 | Human/Mouse |
| 来源宿主 | Rabbit IgG |
| 应用 | W/IP/IF-IC/F/N |
| 使用方法 | WB(1:1000) IP (1:50) F (1:200) IF-IC (1:500) N (1:100) |
| 供应商 | CST |
| 背景 | 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). |
| 存放说明 | -20C |
| 计算分子量 | 175 |
Confocal immunofluorescent analysis of A549 (left) and CAMA-1 (right) cells using EGF Receptor (D1D4J) XP®Rabbit mAb (Neutralizing) (green). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye). | |
Flow cytometric analysis of fixed, unpermeabilized (left) and live (right) CAMA-1 cells (blue), A549 cells (red), and A-431 cells (green) using EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing). Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. | |
High-content analysis (above) of serum-starved A549 cells that were pre-incubated (1 hr) with increasing concentrations of EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) and subsequently stimulated with hEGF #8916 (15 min, 100 ng/ml). Cells were then fixed and stained for activated EGF Receptor (Phospho-EGFR Tyr1068). Data capture was performed using an ArrayScan™ VTI. Quantification of phosphorylated EGFR suggests an optimal working concentration of 5.0 μg/ml for EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) when blocking hEGF-mediated activity. Confocal immunofluorescent analysis (below) of serum-starved A549 cells that were pre-incubated (1 hr) with either 0.1 μg/ml (lower left) or 5.0 μg/ml (lower right) of EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) and subsequently stimulated with hEGF #8916 (15 min, 100 ng/ml). Cells were then fixed and stained for activated EGF Receptor (Phospho-EGFR Tyr1068) (green) and EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (red) to demonstrate the neutralizing ability of the antibody treatment. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Western blot analysis of extracts from A549, mIMCD-3, and CAMA-1 cells using EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (upper), or β-Actin (D6A8) Rabbit mAb #8457 (lower). | |
Western blot analysis of extracts from serum-starved A549 cells that were pre-incubated (1 hr, 5 μg/ml) with EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (+), or vehicle control (-), and subsequently stimulated with hEGF #8916 (15 min, 100 ng/ml; +), using Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb #3777 (upper), EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (middle), or β-actin (D6A8) Rabbit mAb #8457 (lower). | |
ELISA analysis of extracts from serum-starved A549 cells that were pre-incubated (1 hr, 5 μg/ml) with either vehicle control or EGF Receptor (D1D4J) XP® Rabbit mAb (Neutralizing) (antibody-treated). The extracts were subsequently left unstimulated (red) or stimulated with hEGF #8916 (15 min, 100 ng/ml; blue), using PathScan® Phospho-EGF Receptor (panTyr) Sandwhich ELISA Kit #7911. |
