MA5-14864

antibody from Invitrogen Antibodies

Targeting: ERBB3 HER3, LCCS2

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-14864

Provider

Invitrogen Antibodies

Product name

Phospho-ErbB3 (Tyr1197) Monoclonal Antibody (J.996.0)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Antibody clone number

J.996.0

Concentration

32 µg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of SK-BR-3 (Lane 1), SK-BR-3 treated with Heregulin (50 ng/mL for 30 minutes) (Lane 2), SK-BR-3 treated with Gefitinib followed by Heregulin (1 uM for 16 hours, 50 ng/mL for 30 minutes) (Lane 3), SK-BR-3 treated with Afatinib followed by Heregulin (0.5 uM for 6 hours, 50 ng/mL for 30 minutes) (Lane 4), MCF7 (Lane 5), MCF7 treated with Heregulin (50 ng/mL for 30 minutes) (Lane 6), MCF7 treated with Afatinib followed by Heregulin (0.5 uM for 6 hours, 50 ng/mL for 30 minutes) (Lane 7), MDA-MB-231 (Lane 8), MDA-MB-231 treated with Heregulin (50 ng/mL for 30 minutes) (Lane 9) and MDA-MB-231 treated with Afatinib followed by Heregulin (0.5 uM for 6 hours, 50 ng/mL for 30 minutes) (Lane 10). The blots were probed with Anti-Phospho-ErbB3 (Tyr1197) Rabbit Monoclonal Antibody (Product # MA5-14864, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 185 kDa band corresponding to Phospho-ErbB3 (Tyr1197) was detected and observed to increase upon Heregulin treatment across cell lines tested, but not in MDA-MB-231, which is a negative control for ErbB3. Pre-treatment with ErbB3-antagonist, Afatinib, resulted in inhibition of Phospho-ErbB3 in SK-BR-3 and MCF7 cell lines. However, Gefitinib, a non-ErbB3 an