- PA5-16305 - Invitrogen Antibodies ERBB2 antibody

Submitted references A Microfluidic Immunostaining System Enables Quality Assured and Standardized Immunohistochemical Biomarker Analysis.

Kwon S, Cho CH, Kwon Y, Lee ES, Park JK
Scientific reports 2017 Apr 5;7:45968

The use of mouse models of breast cancer and quantitative image analysis to evaluate hormone receptor antigenicity after microwave-assisted formalin fixation.

Engelberg JA, Giberson RT, Young LJ, Hubbard NE, Cardiff RD
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society 2014 May;62(5):319-34

The mucin MUC4 and its membrane partner ErbB2 regulate biological properties of human CAPAN-2 pancreatic cancer cells via different signalling pathways.

Jonckheere N, Skrypek N, Merlin J, Dessein AF, Dumont P, Leteurtre E, Harris A, Desseyn JL, Susini C, Frénois F, Van Seuningen I
PloS one 2012;7(2):e32232

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF7 (Lane 1), A431 (Lane 2), Raji (Lane 3), HEK-MSR (Lane 4), and T47D (Lane 5). The blots were probed with Anti HER-2 Rabbit Polyclonal Antibody (Product # PA5-16305, 5-10 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (Product # G-21234, 1:5000 dilution). A 136 kDa band corresponding to HER-2 was observed across cell lines tested. An additional 62 kDa band was observed in MCF7 and A431. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with PierceTM Power Blotter System (Product # 22834). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

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 (1), T-47D (2) and MDA-MB-231 (3). The blot was probed with anti-ErbB2 Rabbit Polyclonal Antibody (Product # PA5-16305, 1 µg/mL) 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 ErbB2 was observed in SK-BR-3 and moderate in T-47D where as it was not observed in MDA-MB-231 which is an ErbB2 negative cell line. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane using the wet transfer system. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human breast cancer stained with c-erbB2 antibody using peroxidase-conjugate and AEC chromogen. Note membrane staining of tumor cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of HER-2 using HER-2 Polyclonal Antibody (Product # PA5-16305) on denatured Human SKBR3 Cells.

PA5-16305

Antibody data