- MA5-14488 - Invitrogen Antibodies ERBB2 antibody

Submitted references The clinicopathological and prognostic significance of CD24, CD44, CD133, ALDH1 expressions in invasive ductal carcinoma of the breast: CD44/CD24 expression in breast cancer.

Kapucuoğlu N, Bozkurt KK, Başpınar Ş, Koçer M, Eroğlu HE, Akdeniz R, Akçil M
Pathology, research and practice 2015 Oct;211(10):740-7

Diagnostic and therapeutic strategy and the most efficient prognostic factors of breast malignant fibrous histiocytoma.

Qiu SQ, Wei XL, Huang WH, Wu MY, Qin YS, Li YK, Zhang GJ
Scientific reports 2013;3:2529

STAT3, a Poor Survival Predicator, Is Associated with Lymph Node Metastasis from Breast Cancer.

Chen Y, Wang J, Wang X, Liu X, Li H, Lv Q, Zhu J, Wei B, Tang Y
Journal of breast cancer 2013 Mar;16(1):40-9

Quantitation of fixative-induced morphologic and antigenic variation in mouse and human breast cancers.

Cardiff RD, Hubbard NE, Engelberg JA, Munn RJ, Miller CH, Walls JE, Chen JQ, Velásquez-García HA, Galvez JJ, Bell KJ, Beckett LA, Li YJ, Borowsky AD
Laboratory investigation; a journal of technical methods and pathology 2013 Apr;93(4):480-97

The prognostic value of breast cancer resistance protein (BCRB/ABCG2) expression in breast carcinomas.

Omran OM
Journal of environmental pathology, toxicology and oncology : official organ of the International Society for Environmental Toxicology and Cancer 2012;31(4):367-76

The prognostic value of breast cancer resistance protein (BCRB/ABCG2) expression in breast carcinomas.

Omran OM
Journal of environmental pathology, toxicology and oncology : official organ of the International Society for Environmental Toxicology and Cancer 2012;31(4):367-76

Combinatorial treatment of mammospheres with trastuzumab and salinomycin efficiently targets HER2-positive cancer cells and cancer stem cells.

Oak PS, Kopp F, Thakur C, Ellwart JW, Rapp UR, Ullrich A, Wagner E, Knyazev P, Roidl A
International journal of cancer 2012 Dec 15;131(12):2808-19

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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 SK-OV-3 (Lane 1), T-47D (Lane 2), MCF7 (Lane 3) and Hep G2 (Lane 4). The blots were probed with Anti-HER-2 Rabbit Monoclonal Antibody (Product # MA5-14488, 1:250 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conj µgate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 138 kDa band corresponding to HER-2 was observed across the cell lines tested. An additional non-specific band around 60 kDa was observed in SKOV-3 is . Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), 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 overnight 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: 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 blots were probed with Anti-ErbB2 Mouse Monoclonal Antibody (Product # MA5-14488, 1:250 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 185 kDa band corresponding to ErbB2 was observed in SK-BR-3 and T-47D, but not in MDA-MB-231 which is a negative control for ErbB2. 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: Immunofluorescence analysis of ErbB2 was performed using 90% confluent log phase SK-OV-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ErbB2 (EP1045Y) Rabbit Monoclonal Antibody (Product # MA5-14488) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization in the membrane. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Her2 was performed using 70% confluent log phase SK-BR-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Her2 Rabbit monoclonal Antibody (Product # MA5-14488) at 1:250 dilution in 0.1% BSA and incubated overnight at 4 degree Celsius and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous and cytoplasmic localization. Panel f represents MDAMB-231 cells as negative controls, showing no Her2 staining. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded breast carcinoma stained with rabbit monoclonal anti-HER-2 antibody using peroxidase conjugate and DAB chromogen. Note membrane staining of tumor cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of HER-2 / ErbB2 was done on SK-OV-3 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with HER-2 / ErbB2 Rabbit Monoclonal Antibody (Product # MA5-14488, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10, 000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

MA5-14488

Antibody data