HER2 / ErbB2 Recombinant Rabbit Monoclonal Antibody [PD00-53]
cat.: HA721178
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: IHC-P, WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: PD00-53
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 138 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human ErbB2/ HER2 aa 500-650.
Positive control: Human breast carcinoma tissue, SK-Br-3 cell lysates, MCF-7, SK-Br-3.
Subcellular location: Cell membrane, Nucleus, Cytoplasm.
Recommended Dilutions:
  IHC-P
  WB
  IF-Cell
  FC

1:2,000
1:1,000
1:50
1:1,000
Uniprot #: SwissProt: P04626 Human
Alternative names: erb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog C erb B2/neu protein CD340 CD340 antigen Cerb B2/neu protein CerbB2 Erb b2 receptor tyrosine kinase 2 ErbB-2 proto-oncogene ERBB2 ERBB2_HUMAN HER 2 HER 2/NEU HER2 Herstatin Human epidermal growth factor receptor 2 Metastatic lymph node gene 19 protein MLN 19 MLN19 NEU NEU proto oncogene Neuro/glioblastoma derived oncogene homolog Neuroblastoma/glioblastoma derived oncogene homolog NGL p185erbB2 Proto-oncogene c-ErbB-2 Proto-oncogene Neu Receptor tyrosine-protein kinase erbB-2 TKR1 Tyrosine kinase type cell surface receptor HER2 Tyrosine kinase-type cell surface receptor HER2 V erb b2 avian erythroblastic leukemia viral oncogene homolog 2 (neuro/glioblastoma derived oncogene homolog) V erb b2 avian erythroblastic leukemia viral oncogene homolog 2 V erb b2 avian erythroblastic leukemia viral oncoprotein 2 V erb b2 erythrobl......
Images
HA721178_1.jpg Fig1: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-HER2 / ErbB2 antibody (HA721178) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721178) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721178_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-HER2 / ErbB2 antibody (HA721178) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721178) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721178_3.jpg Fig3: Western blot analysis of HER2 / ErbB2 on SK-Br-3 cell lysates with Rabbit anti-HER2 / ErbB2 antibody (HA721178) at 1/1,000 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 138 kDa
Observed band size: 250 kDa

Exposure time: 2 minutes;

6% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721178) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
HA721178_4.jpg Fig4: Immunocytochemistry analysis of MCF-7 cells labeling HER2 / ErbB2 with Rabbit anti-HER2 / ErbB2 antibody (HA721178) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-HER2 / ErbB2 antibody (HA721178) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
HA721178_5.jpg Fig5: Immunocytochemistry analysis of SK-Br-3 cells labeling HER2 / ErbB2 with Rabbit anti-HER2 / ErbB2 antibody (HA721178) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-HER2 / ErbB2 antibody (HA721178) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
HA721178_6.jpg Fig6: Flow cytometric analysis of SK-Br-3 cells labeling HER2 / ErbB2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721178, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.