700635

antibody from Invitrogen Antibodies

Targeting: ERBB2 CD340, HER-2, HER2, NEU, NGL

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

700635

Provider

Invitrogen Antibodies

Product name

ErbB2 (HER-2) Recombinant Rabbit Monoclonal Antibody (40H87L57)

Antibody type

Monoclonal

Antigen

Other

Description

This antibody is predicted to react with canine, chicken, feline, mouse, rat, Rhesus monkey and Xenopus based on sequence homology.

Antibody clone number

40H87L57

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HER-2 in A431 cell lysate, either untreated (lane 4) or serum-starved overnight before addition of 200 ng/mL EGF for 30 min (lanes 1-3) using a HER-2 recombinant rabbit monoclonal antibody (Product # 700635) at a dilution of 2 µg/mL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-ErbB2 (HER-2) Recombinant Rabbit Monoclonal Antibody (40H87L57) (Product # 700635) and a ~180kDa band corresponding to Receptor tyrosine-protein kinase erbB-2 was observed across cell lines tested . Whole cell extracts (50 µg lysate) of SK-BR-3 (Lane 1), MCF7 (Lane 2), SK-O-V3 (Lane 3), Hep G2 (Lane 4), HEK-293 (Lane 5), T-47D (Lane 6), THP-1 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (2 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).Increased expression observed in SK-BR-3, SKOV-3 and HepG2 compared to other cell lines.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Receptor tyrosine-protein kinase erbB-2 was performed using 70% confluent log phase SK-BR-3 and T-47D cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with ErbB2 (HER-2) Recombinant Rabbit Monoclonal Antibody (40H87L57) (Product # 700635) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cell membrane, endosome localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.