Human Phospho-ErbB2/Her2 (Y1248) Antibody

Detection of Human Phospho-ErbB2/Her2 (Y1248) by Western Blot. Western blot shows lysates of MDA-MB-468 human breast cancer cell line and A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 1 mM Pervanadate (PV) for 10 minutes or with 10 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 0.25 µg/mL of Rabbit Anti-Human Phospho-ErbB2/Her2 (Y1248) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1768) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-ErbB2/Her2 (Y1248) at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

ErbB2/Her2 in Human Breast Cancer Tissue. ErbB2/Her2 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human Phospho-ErbB2/Her2 (Y1248) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1768) at 0.3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Human Phospho-ErbB2/Her2 (Y1248) by Simple Western^TM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-ErbB2/Her2 (Y1248) at approximately 265 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-ErbB2/Her2 (Y1248) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1768). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

Detection of Human ErbB2/Her2 by Western Blot Antitumor effects of lapatinib and PD168393 in the ErbB2 positive lung cancer cell line, Calu3 and the ErbB2 positive breast cancer cell line, SkBr3 after 72 hour treatment.A. Cellular proliferation of Calu3 and SkBr3 cells treated with different concentrations of lapatinib and PD168393 as determined by the MTS assay. B. Lapatinib and PD168393 induce cellular apoptosis in Calu3 and SkBr3 cells. Representative Annexin V/propidium iodide flow cytometry; the numbers represent percentage of cells in the appropriate quadrant. C. Quantification of apoptosis. D. Lapatinib induces cell cycle arrest in Calu3 cells as detected by BrdU and 7-ADD assay. R1-G1 phase; R2-G2 phase; R3-S phase; R4-G0 phase. E. Dose response of lapatinib and PD168393 on phosphorylation of ErbB2, AKT and ERK in Calu3 and SkBr3 cells in response to EGF treatment. p-ErbB2: phosphorylated ErbB2; t-ErbB2: total-ErbB2; p-AKT: phosphorylated AKT; t-AKT: total AKT; p-ERK: phosphorylated ERK; t-ERK: total ERK. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0106349), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human ErbB2/Her2 by Western Blot Downregulation of PHLDA1 expression upon ErbB2/EGFR inhibition.Western blot analysis following treatment with lapatinib, erlotinib and CL387,785 at 1 µm for 6, 24 and 48 hours in ErbB2-positive SkBr3 and EGFR-mutated H1975, HCC827. DMSO treatment was 48 hours. Different cell lines were used with the relevant inhibitor capable of blockade of oncogenic signaling. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0106349), licensed under a CC-BY license. Not internally tested by R&D Systems.