anti-p-egfr(y992)  (Cell Signaling Technology Inc)

Journal: ACS Omega

Article Title: Novel Adamantanyl-Based Thiadiazolyl Pyrazoles Targeting EGFR in Triple-Negative Breast Cancer

doi: 10.1021/acsomega.6b00251

Figure Lengend Snippet: Cheminformatics and surface plasmon resonance (SPR) analysis predicts the interaction of APP with the EGFR protein. (A) Predicted molecular interactions between EGFR and APP: (i) Template crystal structure of EGFR (gray cartoon) in complex with a hydrophobic kinase inhibitor (cyan cartoon). (ii) The predicted binding mode of APP shows a major shape overlap with the co-crystallized ligand. Main interaction centers are highlighted as thin sticks and include Leu-718, Val-726, and Lys-745, which form hydrogen bonds to the ligand (yellow dots). (B) The sensorgrams obtained by SPR analysis of APP with the EGFR protein subunit. The EGFR protein subunit was immobilized onto the surface of a CM5 sensor chip. A solution of APP at variable concentrations was injected to generate the results of binding responses (RU) recorded as a function of time (s). The results were analyzed using BIA evaluation 3.1. (C) Western blot analysis was performed to evaluate the effect of APP on EGFR phosphorylation (at Y992, Y1068, Y1086, Y1148, and Y1173) in BT549 cells. Soluble whole cell extracts were run on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted as described in . β-Actin was used as input control for cell lysate. The sizes of the detected protein bands in kilodaltons are shown on the left side .

Article Snippet: Rabbit anti-p-EGFR (Y992) and rabbit anti-p-CDK2 antibodies were obtained from Cell Signaling.

Techniques: SPR Assay, Binding Assay, Injection, Western Blot, Phospho-proteomics, Polyacrylamide Gel Electrophoresis, SDS Page, Control

Journal: Molecular medicine reports

Article Title: Inhibitory effects of tetrandrine on epidermal growth factor-induced invasion and migration in HT29 human colorectal adenocarcinoma cells.

doi: 10.3892/mmr.2015.4635

Figure Lengend Snippet: Figure 5. Effects of tetrandrine on EGFR signaling in EGF‑induced HT29 cells. Prior to 100 ng/ml EGF exposure, the cells were treated with or without 0.5, 1 and 2 µM of tetrandrine. After 48 h, the cell lysates were harvested and subjected to western blot analysis. The protein expression levels of p‑EGFR (Y845), p‑EGFR (Y992), p‑EGFR (Y1068) and EGFR were determined and adjusted for equivalent loading using the Actin antibody. EGF, epidermal growth factor; p‑EGFR, phosphorylated EGFR.

Article Snippet: Anti‐p‐EGFR (Y845) (2231; 1:1,000), anti-p-EGFR (Y992) (2235; 1:1,000), anti-p-EGFR (Y1068) (2234; 1:1,000), anti-p-phosphoinositide-dependent kinase-1 (PDK1; 3031; 1:1,000), anti-p-PI3K (4228; 1:1,000), anti-p-AKT (S308) (13038; 1:1,000), anti-p-AKT (S473) (4060; 1:1,000), anti-p-ERK (Thr202/Tyr204) (4370; 1:1,000), anti-p-c-Jun N-terminal kinase (JNK; Thr183/Tyr185) (4668; 1:1,000) and anti-p-p38 (Thr180/Tyr182) (4511; 1:1,000) antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).

Techniques: Western Blot, Expressing