MA5-13105

antibody from Invitrogen Antibodies

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

Provider product page for MA5-13105

Western blot

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [131]
Comments [0]
Validations
Western blot [3]
Immunohistochemistry [2]
Flow cytometry [2]
Other assay [9]

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Product number: MA5-13105 - Provider product page
Provider: Invitrogen Antibodies
Product name: ErbB2 (HER-2) Monoclonal Antibody (e2-4001)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: MA5-13105 targets HER-2 in IHC, ICC/IF, IP, FACS and WB applications and shows reactivity with human, mouse, and rat samples.
Antibody clone number: e2-4001
Concentration: 400 µg/mL

ERBB2 protein structure - MA5-13105 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 # MA5-13105) on SKBR3 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 # MA5-13105, 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 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 # MA5-13105, 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: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of HER-2 using HER-2 Monoclonal Antibody (Product # MA5-13105) on denatured Human SKBR3 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 Nf1 deficiency induces ER + /PR + mammary tumors. We observed mammary tumors with diverse mammary histology in G0 - G2 animals. a Nf1 females and b Nf1 males developed mammary adenocarcinomas with mixed histology including acinar, cystic, papillary, ductal, and solid features. All images were taken at 100X magnification. c Immunostaining of Nf1 mammary tumors ( rNf1 #6) for estrogen receptor, progesterone receptor, HER2, Ki67, and pan-cytokeratin (AE1/AE3). All immunostaining images were taken at 200X magnification

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 T-DM1 increases VSVDelta51 activity in a panel of HER2-expressing breast and ovarian cancer lines. a A panel of human breast and ovarian carcinoma lines were lysed and subject to western blotting for human HER2. b Mouse breast carcinoma 4T1 cells, and a panel of human cancer cell lines were subject to extracellular staining with Herceptin(r), followed by goat anti-human IgG-PE. PE signal ( y axis) was analyzed by flow cytometry, median MFI are shown; n = 3 +- SD. c 786-0 and JIMT1 cells were pretreated with T-DM1 for 4 h, washed and infected with VSVDelta51 (MOI 0.05 and 0.01, respectively) and plaque assay performed 45 h later ( n = 3; mean +- SEM; one-way ANOVA with Dunnett's multiple comparisons test compared to 0 µg/mL T-DM1 for each cell line). d Indicated cell lines were pretreated with either 100 nM colchicine, 100 mug/mL T-DM1 or trastuzumab, or mock, for 3 h, then washed and infected with VSVDelta51 (MOI 0.01 for JIMT1 and MOI 0.001 for others). 1 h later, wells were overlayed with agarose, and plaques visualized with Coomassie blue 48 h later. Diameter of 30 plaques per well were graphed (mean +- SEM; one-way ANOVA with Dunnett's multiple comparisons test compared to Mock for each cell line). e JIMT1 cells were plated on glass coverslips, treated with 100 µg/mL T-DM1 or trastuzumab, or mock-treated, then stained for beta-tubulin and DAPI 24 h later. Objective = x63, scale bar = 50 um, n = 3. White arrow indicates polynuclear cells. f Cell lines in

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 T-DM1 increases oncolytic activity of VSVDelta51 in vivo and in human patient samples. a BALB/c nude mice were implanted with 1 x 10 7 JIMT1 cells. Once tumors were ~100 mm 3 , mice were treated weekly four times (dotted lines) with the regimen of VSVDelta51-Fluc (1E8 pfu) or PBS (intratumorally) followed by 10 mg/kg T-DM1 (Kadcyla(r)) or PBS 4 h later (intravenously). Survival was monitored over time ( n = 16-20 mice per group as indicated, pooled over three independent experiments) and was observed to be significantly reduced with the combination of VSVDelta51 + T-DM1 compared to either monotherapies ( p = 0.039 compared to Kadcyla; p = 0.004 compared to VSV; p < 0.0001 compared to PBS, Gehan-Breslow-Wilcoxon test). b Ascites fluid cells derived from three different human ovarian cancer patients were each lysed and blotted for HER2 or actin as a loading control. c Mouse breast carcinoma 4T1 cells, and the ascites fluid-derived cell lines were subject to extracellular staining and flow cytometry. Cells were stained with Herceptin(r) (trastuzumab, 1:1000), followed by goat anti-human IgG-PE (1:100). PE signal ( y axis) was analyzed by flow cytometry, median MFI are shown; n = 3 +- SD. d AF2028 and AF2068 were plated on glass coverslips, treated with 100 µg/mL T-DM1 or trastuzumab, then fixed and stained for beta-tubulin and DAPI 24 h later. Objective = x63, scale bar = 50 um, n = 3. White arrow indicates polynuclear cells. e The ascites fluid-derived cell lines w

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 MLH1 loss in ER + , nominally HER2 - breast cancer cells upregulates membrane-bound HER2. A Western blots demonstrating increase in pHER2 and downstream signaling in sh MLH1 MCF7 cells treated with fulvestrant relative to sh Luc cells. Quantification of four independent replicates conducted through ImageJ in accompanying dot plots. Validation in T47D cells in Fig. S3A . Immunofluorescent staining for HER2 in MCF7 sh Luc and sh MLH1 cells in vitro ( B ), in MCF7 sh Luc and sh MLH1 xenograft tumors ( C ), and in WHIM20, PMS2 mutant, ER + /HER2 - PDX tumors ( D ), grown with or without fulvestrant. Accompanying quantification presented as strip charts. Three independent experiments or tumors from each group were quantified. Two-sided Student's t test determined p values. Supporting data from FACS analysis are presented in Fig. S3B, C . Scale bars represent 50 u. Source data for all figures available with paper.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 MLH1 regulates protein trafficking of HER2. A Gene set enrichment analysis of RNAseq data comparing isogenic sh Luc and sh MLH1 MCF7 cells after treatment with fulvestrant for 4 days. P values were generated using DESeq2 R package and adjusted for multiple comparison using Benjamini-Hochberg. Comparable RPPA data analysis in Fig. S4A . Raw read counts available as supplementary data. B Co-immunofluorescence for HER2 and lysosomal marker, LAMP1 (orange arrows) at baseline and after 18, 36, and 54 h of fulvestrant treatment. Green arrows indicate HER2 that is not colocalized with LAMP1, and red arrows indicate LAMP1 positivity alone. Validation in T47D cells in Fig. S4B . C Immunofluorescence staining for HER2 in MCF7 sh Luc and sh MLH1 cells treated with vehicle and 36 h of fulvestrant alone or a combination of fulvestrant and chloroquine, an autophagy inhibitor. For sh Luc vs. sh MLH1 vehicle, p = 0.0006. Validation in T47D cells in Fig. S4C . Quantification ( D ) and representative photomicrographs ( E ) from 36 h of live cell tracking of colocalization of HER2 and LC3 (orange arrows), a marker of autophagosomes, in MCF7 sh Luc and sh MLH1 cells treated with vehicle or fulvestrant. Green arrows indicate HER2 + LC3 - cells. Cells were tracked after administration of fulvestrant. All quantification is of three independent biological replicates conducted through ImageJ and is represented as strip charts. Two-sided Student's t test determined all p values. For sh Luc vehi

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 10 Immunohistochemical staining of the xenograft tumor sections (400X). Dark brown color was observed as positive staining. Tissues from xenograft tumors were negative for ER (A) , PR (B) and HER-2 (C) , and were positive for P63 (D) , CK5/6 (E) , and alpha-SMA (F) localized in the cell membrane.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 The rare-cell sc WB measures 12 unique protein targets in single cells from three cancer subtypes. ( a ) Fluorescence micrographs and intensity plots from rare-cell sc WB handling and analysis of healthy blood samples, each spiked with a cancer subtype: EGFR+(BT-20), ER+(MCF7) and HER2+(SK-BR-3). Negative controls include analysis of WBCs only and blank microwells (that is, devoid of a cancer cell). Protein panel comprises the following: control and housekeeping proteins (GAPDH and beta-tubulin), oncoproteins (HER2, ER and EGFR), signalling proteins (ERK, eIF4E and mTOR), common CTC classifiers (EpCAM, panCK and CK8) and a WBC indicator (CD45). Scale bar, 50 mum. ( b ) Comparative protein expression for each cancer cell (BT-20: n =27; SK-BR-3: n =27; MCF7: n =35), with mean and CV noted for each marker. Protein expression is graphed using a log-scale. Ranked oncoprotein expression for each cell line agrees with cancer subtype. Less than 1% of total protein signal is attributable to CD45.