ErbB2/Her2 [p Tyr1139] Antibody (JE60-90)

Western Blot: ErbB2/Her2 [p Tyr1139] Antibody (JE60-90) [NBP3-32768] - Western blot analysis of ErbB2/Her2 on different lysates with Rabbit anti-ErbB2/Her2 antibody (NBP3-32768) at 1/1,000 dilution. Lane 1: A431 cell lysate Lane 2: A431 treated with 100ng/mL EGF for 10 minutes cell lysate Lane 3: A431 treated with 100ng/mL EGF for 10 minutes cell lysate, then the membrane treated with pp for 1 hour Lane 4: SK-Br-3 cell lysate Lane 5: SK-Br-3 starved for 4 hours add 200ng/mL EGF for 15 minutes cell lysate Lysates/proteins at 20 ug/Lane. Predicted band size: 138 kDa Observed band size: 180 kDa Exposure time: 10 seconds; ECL; 4-20% 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 (NBP3-32768) at 1/1,000 dilution was used in 5% NFDM/TBST at 4C overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry/ Immunofluorescence: ErbB2/Her2 [p Tyr1139] Antibody (JE60-90) [NBP3-32768] - Immunocytochemistry analysis of A431 cells treated with or without 100ng/mL EGF for 10 minutes labeling ErbB2/Her2 with Rabbit anti-ErbB2/Her2 antibody (NBP3-32768) at 1/5,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ErbB2/Her2 antibody (NBP3-32768) at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.

Flow Cytometry: ErbB2/Her2 [p Tyr1139] Antibody (JE60-90) [NBP3-32768] - Flow cytometric analysis of A431 cells treated with or without 100ng/mL EGF for 10 minutes labeling ErbB2/Her2. Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32768, 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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

• Reactivity: Hu
• Applications: WB, Flow, ICC/IF
• Clone: JE60-90
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

ErbB2/Her2 [p Tyr1139] Antibody (JE60-90) Summary

• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Tyr1139 of Human ErbB2/Her2. (Uniprot: P04626)
• Modification: p Tyr1139
• Localization: Cell membrane, Cell projection, ruffle membrane; Early endosome, Cytoplasm, perinuclear region, Nucleus.
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: ERBB2
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:5000
  • Western Blot 1:1000
• Theoretical MW: 138 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for ErbB2/Her2 [p Tyr1139] Antibody (JE60-90)

• CD340 antigen
• CD340
• c-erb B2/neu protein
• EC 2.7.10
• EGFR2
• ErbB2
• HER2
• HER-2
• HER2EC 2.7.10.1
• herstatin
• Metastatic lymph node gene 19 protein
• MLN 19
• MLN19
• Neu Oncogene
• NEUHER-2/neu
• neuroblastoma/glioblastoma derived oncogene homolog
• NGL
• NGLTKR1
• p185erbB2
• Proto-oncogene c-ErbB-2
• Proto-oncogene Neu
• receptor tyrosine-protein kinase erbB-2
• TKR1
• 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 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastomaderived oncogene homolog (avian)

Background

ErbB2/HER2 is one of the four members of the ErbB receptor family of transmembrane receptor-like tyrosine kinases (1). The kinase activity of ErbB2 can be activated without ligand if it is overexpressed, and by association with other ErbB proteins (2). Overexpression of ErbB2 is detected in almost 40% of human breast cancers (3). Binding of c-Cbl ubiquitin ligase to Tyr1112 of ErbB2 leads to poly-ubiquitination of ErbB2 and enhances its degradation (4). ErbB2 is one of the major targets for the treatment of breast cancer and other carcinomas.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

FAQs for ErbB2/Her2 Antibody (NBP3-32768). (Showing 1 - 1 of 1 FAQ).

1. I am interested in getting antibodies for HER that would work in flow cytometry. Could you recommend some choices?
   • A list of our ErbB2 antibodies validated for use in flow cytometry can be found here /productsearch/HER2#fq=common_name%3A%22ErbB2%2FHER2%22&fq=category%3A%22Primary%20Antibodies%22&fq=applications%3A%22Flow%20Cytometry%22

Bioinformatics

• Gene Symbol: ERBB2