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

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

40H87L57

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

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.