MA1-35720

antibody from Invitrogen Antibodies

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

Provider product page for MA1-35720

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [47]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [1]
Other assay [18]

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Product number: MA1-35720 - Provider product page
Provider: Invitrogen Antibodies
Product name: ErbB2 (HER-2) Monoclonal Antibody (CB11)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: CB11
Vial size: 1 mL
Concentration: Conc. Not Determined
Storage: 4° C

ERBB2 protein structure - MA1-35720 shown in red.

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 # MA1-35720, 1:1000 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 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 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 blot was probed with Anti- ErbB2 Mouse Monoclonal Antibody (Product # MA1-35720, 1:1000 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 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 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)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 1 Loss of HER2 prevents the formation of membrane protrusion in HER2KD-SKBR3 cells. A) Examination of relative expression of HER2 by QPCR. Bars represent mean +- SEM for 3 separate experiments. B) Immunoblot analysis from control and HER2KD-SKBR3 cells. C) Time course of pHER2 and pAKT levels in control (black bars) and HER2KD-SKBR3 (gray bars) cells in response to EGF. Cells were serum-starved for 16 hours, treated with growth factors and harvested at times listed on graphs. Each bar represents the mean +- SEM of 3 separate experiments. * represents p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 5 HER2 signaling prevents HER2 internalization. A) Con-focal images of immunofluorescence for HER2 (green) and EGFR (red) in control (top row) and HER2KD-SKBR3 (bottom row) cells at baseline in serum-containing media. White arrow points to internalized HER2. Exposure times are longer in HER2KD cells to compensate for reduced HER2 levels. B) Con-focal images of immunofluorescence for HER2 (green) and EGFR (red) in control (top row) and HER2KD-SKBR3 (bottom row) cells exposed EGF for 2 hrs. Panels on right represent magnifications of boxed areas. White arrow indicates internalized HER2 and EGFR. C) Quantification of cell surface HER2 as detected by isolation of cell-surface biotinylated proteins. Each bar represents the mean +- SEM of 3 separate experiments in cells at baseline and after EGF treatment. Asterisk denotes p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 4 Lapatinib increases intracellular calcium and inhibits membrane protrusions. A) Immunoblot showing HER2 signaling components in SKBR3 cells +- lapatinib ( 2 micromolar for 48-hours). B) Intracellular calcium measurements in control and lapatinib-treated SKBR3 cells. Numbers are mean calcium concentrations estimated by FURA2 measurements. C) Con-focal images of immunofluorescence for NFATc1 (red) and DAPI (blue) in SKBR3 cells treated with lapatinib. D) Con-focal images of immunofluorescence for HER2 (red) and phalloidin (actin, green) in control and lapatinib-treated SKBR3 cells. Insets represent Z-stack images in 2 different orientations. E) Scanning electron microscopy of control and lapatinib-treated SKBR3 cells. Arrows point to larger membrane protrusions. F) Co-IP for HER2 and HSP90 from control and lapatinib-treated SKBR3 cells. Cell lysates were immunoprecipitated with antibodies to HSP90 and blotted for HSP90 or HER2. G) Con-focal images of immunofluorescence for HER2 (red) and HSP90 (green) in control (top row) and lapatinib-treated (bottom row) SKBR3 cells. Insets represent Z-stack images in 2 different orientations and white arrows indicate internalized HER2. Scale bars in C, D and G represent 10mum. Scale bars in E represent 20muM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6. Enhanced interactions between HER2 and MAL2, Ezrin, NHERF1, and PMCA2 in trastuzumab-resistant SKBR3 cells (A) Immunofluorescence staining for cholera toxin B (lipid rafts) in control and trastuzumab-resistant SKBR3 cells. Scale bars represent 10 mum. (B) Immunofluorescence staining for HER2 and MAL2 in control and trastuzumab-resistant SKBR3 cells. Scale bars represent 10 mum. (C) PLA for HER2 and MAL2 in control and trastuzumab-resistant SKBR3 cells also stained for phalloidin. Scale bars represent 10 mum. (D-F) Quantitative results from immunoprecipitation coupled with data-independent acquisition mass spectrometry (DIA-MS) in control and trastuzumab-resistant SKBR3 cells. (D) The DIA-MS Intensity (log 2 ) of HER2 and MAL2 proteins from control and trastuzumab-resistant SKBR3 cells. (E) The DIA-MS Intensity (log 2 ) for all the peptide precursor signals of HER2 in control and resistant cells. (F) The DIA-MS peak groups visualized for quantifying HER2 (VLGSGAFGTVYK) and MAL2 (VTLPAGPDILR). Peaks above and below the middle line denote the MS2 and MS1 ion traces in DIA-MS. (G) MAL2, Ezrin, and NHERF1 mRNA expression in control and trastuzumab-resistant SKBR3 cells as assessed by quantitative RT-PCR (n = 3). (H) PLA for HER2 with Ezrin (left), NHERF1 (middle), and PMCA2 (right) in control and trastuzumab-resistant SKBR3 cells also stained for phalloidin. Boxed portions are amplified at right with co-registration of PLA signal and immunofluorescence for actin (phalloi

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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