MA5-13675

antibody from Invitrogen Antibodies

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

Provider product page for MA5-13675

Western blot

Immunocytochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [75]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [1]
Flow cytometry [3]
Other assay [7]

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Product number: MA5-13675 - Provider product page
Provider: Invitrogen Antibodies
Product name: ErbB2 (HER-2) Monoclonal Antibody (3B5)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: MA5-13675 targets HER-2 in IHC, IP, ICC, IF, FACS and WB applications and shows reactivity with Human, mouse, Non-human primate, and Rat samples.
Antibody clone number: 3B5
Concentration: 0.1 mg/mL

ERBB2 protein structure - MA5-13675 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of HER-2/ErbB2 in 3T3-L1 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a HER-2/ErbB2 monoclonal antibody (Product # MA5-13675) at a dilution of 1:50 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of HER-2/ErbB2 in MDA-MB-453 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a HER-2/ErbB2 monoclonal antibody (Product # MA5-13675) at a dilution of 1:50 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of HER-2/ErbB2 in SK-BR-3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a HER-2/ErbB2 monoclonal antibody (Product # MA5-13675) at a dilution of 1:50 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of HER-2 using HER-2 Monoclonal Antibody (Product # MA5-13675) on denatured Human SKBR3 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of HER-2 using HER-2 Monoclonal Antibody (Product # MA5-13675) on Native Human SKBR3 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 at RA induces miR-4680-3p expression in HEPM cells. (A) Quantitative RT-PCR for the indicated miRs after treatment of HEPM cells with at RA for 1 or 3 days. *** p < 0.001. (B) Quantitative RT-PCR for the indicated genes after treatment of HEPM cells with at RA for 1 or 3 days. * p < 0.05, ** p < 0.01. Each treatment group was compared with a control vehicle group at each indicated day. (C) Immunoblotting for ERBB2, JADE1, phosphorylated ERK1/2 (P-ERK1/2), ERK1/2, phosphorylated mTOR (P-mTOR), mTOR, and GAPDH in HEPM cells treated with 30 muM at RA for 72 h. Representative images from two independent experiments are shown. (D) Immunocytochemical analysis of ERBB2 and JADE1 (green) in HEPM cells treated with 30 muM at RA for 72 h. Nuclei were counterstained with DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 ERBB2 and JADE1 knockdown inhibits cell proliferation in HEPM cells. (A) Quantitative RT-PCR for ERBB2 after treatment with ERBB2 siRNA for 24 h in HEPM cells. *** p < 0.001. (B) Immunoblotting of ERBB2 and GAPDH in HEPM cells treated with an ERBB2 siRNA for 48 h. Representative images from two independent experiments are shown. (C) Quantitative RT-PCR for JADE1 after treatment with JADE1 siRNA for 24 h in HEPM cells. *** p < 0.001. (D) Immunoblotting of JADE1 and GAPDH in HEPM cells treated with JADE1 siRNA for 48 h. Representative images from two independent experiments are shown. (E) Cell proliferation assays in HEPM cells treated with an ERBB2 or JADE1 siRNA for 24, 48, or 72 h. ** p < 0.01, *** p < 0.001. Each treatment group was compared with a control siRNA group at each indicated day. (F) Immunoblotting of CCND1, CDKN1B, and GAPDH in HEPM cells treated with siRNA for ERBB2 or JADE1 for 48 h. Representative images from two independent experiments are shown. (G) ERBB2 expression following overexpression in HEPM cells. ** p < 0.01. (H) JADE1 expression following overexpression in HEPM cells. *** p < 0.001. (I) Cell proliferation assays in HEPM cells overexpressing ERBB2 and/or JADE1 for 24, 48, or 72 h. ** p < 0.01, *** p < 0.001. Each treatment group was compared with control vector (mock) group at each indicated day. ## p < 0.01 vs. mock + at RA at day 3.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Normalization of miR-4680-3p expression restores at RA-induced cell proliferation suppression in HEPM cells. (A) Cell proliferation assays in HEPM cells treated with 30 muM at RA, with/without miR-4680-3p inhibitor, for 24, 48, or 72 h. *** p < 0.001 vs. control inhibitor + vehicle at each indicated day. ## p < 0.01 vs. control inhibitor + at RA at indicated day. (B) BrdU staining (red) in HEPM cells after treatment with 30 muM at RA, with/without miR-4680-3p inhibitor, for 72 h. Nuclei were counterstained with DAPI (blue). (C) Graph shows the quantification of BrdU-positive cells. ** p < 0.01 vs. control inhibitor + vehicle. ## p < 0.01 vs. control inhibitor + at RA. (D) Quantitative RT-PCR for the indicated genes after treatment with at RA, with/without miR-4680-3p inhibitor, for 24 h in HEPM cells. ** p < 0.01 vs. control inhibitor + vehicle. ## p < 0.01 vs. control inhibitor + at RA. (E) Immunoblotting of ERBB2, JADE1, phosphorylated ERK1/2 (P-ERK1/2), ERK1/2, CCND1, CDKN1B, and GAPDH in HEPM cells treated with 30 muM at RA, with/without miR-4680-3p inhibitor, for 72 h. Representative images from two independent experiments are shown.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Infection schematic and characterization of Control, HER2ic, and HER2ma virus-like particle (VLP) vaccines. In ( A ), the schematic of recombinant baculoviruses (rBVs) used for the generation of VLPs is shown. In ( B - D ), sucrose-density purified VLPs were negatively stained and investigated using a transmission electron microscope (FEI Tecnai G2 200 kV, FEI, Hillsboro, OR, USA). All types of VLPs (( B ) the control, ( C ) HER2ic, ( D ) HER2ma) appeared spherical. In ( E ), the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis results of the VLP preparations are shown. Trastuzumab, a monoclonal anti-HER2 antibody (3B5), and a monoclonal rabbit antihuman HIV-1 Gag antibody were used to probe truncated HER2, full-length human HER2 protein, and the HIV-1 Gag matrix protein, respectively.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Antibody responses upon immunization with Control, HER2ic, and HER2ma VLPs. Mice were immunized with Control, HER2ic, or HER2ma VLPs in a prime-boost regimen, in a nonadjuvant, or in combination with Poly (I:C) or AddaVax. ( A ) Serum antibodies of vaccinated mice were investigated in an ELISA against recombinant HER2 protein. Results are shown as mean +- SEM (standard error of mean) of each vaccination group ( n = 5). In ( B ), IgG1:IgG2a:IgG2b subtype ratios of induced anti-HER2 antibodies elicited by the indicated vaccinations are shown.