MA5-14057

antibody from Invitrogen Antibodies

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

Provider product page for MA5-14057

Western blot

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [141]
Comments [0]
Validations
Western blot [4]
Immunohistochemistry [2]
Flow cytometry [3]
Other assay [3]

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Product number: MA5-14057 - Provider product page
Provider: Invitrogen Antibodies
Product name: ErbB2 (HER-2) Antibody Cocktail
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: MA5-14057 targets HER-2 in IHC (P), IP, ICC/IF, FACS and WB applications and shows reactivity with Human samples.
Antibody clone number: e2-4001, 3B5
Concentration: 150 µg/mL

ERBB2 protein structure - MA5-14057 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of HER-2 using HER-2 Monoclonal Antibody (Product # MA5-14057) on LS174T Cells.

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 Mouse Monoclonal Antibody (Product # MA5-14057, 1 µg/mL) 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 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: Western blot analysis of ERBB2 was performed with 10 µg of A549 cells transfected with Transfection Reagent alone (Lane 1), 100nM Non-Targeting control siRNA (Lane 2), or 100nM siRNA against ERBB2 (Lane 3). Proteins were resolved using a NuPAGE® Novex 4-12% Bis-Tris Gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), and a protein size ladder. Proteins were wet transferred to a Pierce Nitrocellulose Membrane (Product # 88025) OR Pierce PVDF Membrane (Product # 88518) and blocked with Pierce Starting Block T20 (PBS) Blocking Buffer (Product # 37539) for 1 hour at room temperature. ERBB2 was detected at ~ 185 kDa using ERBB2 Mouse monoclonal antibody (Product # MA5-14057) diluted in Pierce Starting Block T20 (PBS) Blocking Buffer 4°C overnight on a rocking platform. Pierce Goat Anti-Mouse (Product # 31437) HRP-Conjugated Antibodies at a 1:2500 dilution were used and chemiluminescent detection was performed using Pierce Supersignal West Dura Maximum Sensitivity Substrate (Product # 37071). Relative density of the bands normalized to GAPDH (36 kDa). ERBB2 Antibody (Product # MA5-14057) confirms silencing of ERBB2 expression.

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 Mouse Monoclonal Antibody (Product # MA5-14057, 1 µg/mL) 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 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: 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 # MA5-14057) 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 PC-12 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 # MA5-14057) 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 # MA5-14057) 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: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5. KLF5 controls cell proliferation in OAC. ( A ) Heatmap of ATAC-seq signal at regions (peak centre +-2.5 kb) with significantly differential accessibility (+-2 x; Q < 0.05) in OE19 cells treated with siERBB2. ( B ) Bar chart of percentage targets and percentage background of de novo discovered motifs at regions closed with siERBB2 treatment in OE19 cells. De novo motifs, called transcription factor with motif match scores (brackets) and p-values are shown. ( C ) Tag density plot of ATAC-seq signal from BO and OAC tissue at regions that demonstrate KLF5 binding in OE19 cells and reduced chromatin accessibility in OE19 cells upon siERBB2 treatment. ( D ) Heatmap of z-score of expression of KLF5 direct target genes in OE19 cells treated with either siNT or siERBB2. Cell cycle related genes are indicated with a black bar. ( E ) Bar chart showing the % relative growth of OE19 cells treated with either siNT or siKLF5. p-Value is shown (n = 3). ( F ) Bar chart showing the % relative growth of OE19-dCas9-KRAB cells treated with either non-targeting guides or guides targeting the KLF5 TSS. p-Value is shown (n = 3). ( G ) Bar chart showing the % relative growth of OE19 cells treated with either siNT or siRNA against the indicated target genes. p-Values are shown (n = 3). ( H ) Kaplan-Meier curves of patient survival across 24 months for high (above median; red) or low (below median; blue) expression of the 97 KLF5 target genes (left), KLF5 (middle) or CLSPN (right). p-Values a

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Expression of ERBB-2 in PPGLs. (A) Weakly positive immunostaining of ERBB-2 in PPGLs. (B) Intermediately positive immunostaining of ERBB-2 in PPGLs. (C) Strongly positive immunostaining of ERBB-2 in PPGLs.