WB
Western blot analysis of Phospho-HER2 / ErbB2 (Y1139) on different lysates with Rabbit anti-Phospho-HER2 / ErbB2 (Y1139) antibody at 1/1000 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 λpp 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 µg/Lane. Exposure time: 10 seconds; 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 at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of A431 cells treated with or without 100ng/mL EGF for 10 minutes labeling Phospho-HER2 / ErbB2 (Y1139) with Rabbit anti-Phospho-HER2 / ErbB2 (Y1139) antibody at 1/5000 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-Phospho-HER2 / ErbB2 (Y1139) antibody at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (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 (594) was used as the secondary antibody at 1/1000 dilution.FC
Flow cytometric analysis of A431 cells (left) and A431 cells treated with 100ng/mL EGF for 10 minutes (right) labeling Phospho-HER2 / ErbB2 (Y1139). Cells were fixed and permeabilized. Then stained with the primary antibody (1/10000) (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 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Phospho-HER2 / ErbB2 (Y1139) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phosphopeptide corresponding to residues surrounding Tyr1139 of Human HER2. |
| Modification | P-Y1139 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFWB |
| WB:1:1000 ICC/IF:1:5000 FC:1:10000 |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | ERBB2 |
|---|---|
| Gene Synonyms | NEU NGL HER2 TKR1 CD340 HER-2 VSCN2 MLN 19 MLN-19 c-ERB2 c-ERB-2 HER-2/neu p185(erbB2) |
| Gene Full Name | erb-b2 receptor tyrosine kinase 2 |
| Gene Summary | This gene encodes a member of the epidermal growth factor (EGF) receptor family of receptor tyrosine kinases. This protein has no ligand binding domain of its own and therefore cannot bind growth factors. However, it does bind tightly to other ligand-bound EGF receptor family members to form a heterodimer, stabilizing ligand binding and enhancing kinase-mediated activation of downstream signalling pathways, such as those involving mitogen-activated protein kinase and phosphatidylinositol-3 kinase. Allelic variations at amino acid positions 654 and 655 of isoform a (positions 624 and 625 of isoform b) have been reported, with the most common allele, Ile654/Ile655, shown here. Amplification and/or overexpression of this gene has been reported in numerous cancers, including breast and ovarian tumors. Alternative splicing results in several additional transcript variants, some encoding different isoforms and others that have not been fully characterized. [provided by RefSeq, Jul 2008] |
| Alternative Names | Verb b2 erythroblastic leukemia viral oncogene homolog 2 neuro/glioblastoma derived oncogene homolog antibody C erb B2/neu protein antibody CD340 antibody CD340 antigen antibody Cerb B2/neu protein antibody CerbB2 antibody Erb b2 receptor tyrosine kinase 2 antibody ErbB-2 proto-oncogene antibody ERBB2 antibody ERBB2_HUMAN antibody Verb b2 erythroblastic leukemia viral oncogene homolog 2 neuro/glioblastoma derived oncogene homolog antibody C erb B2/neu protein antibody CD340 antibody CD340 antigen antibody Cerb B2/neu protein antibody CerbB2 antibody Erb b2 receptor tyrosine kinase 2 antibody ErbB-2 proto-oncogene antibody ERBB2 antibody ERBB2_HUMAN antibody HER 2 antibody HER 2/NEU antibody HER2 antibody Herstatin antibody Human epidermal growth factor receptor 2 antibody Metastatic lymph node gene 19 protein antibody MLN 19 antibody MLN19 antibody NEU antibody NEU proto oncogene antibody Neuro/glioblastoma derived oncogene homolog antibody Neuroblastoma/glioblastoma derived oncogene homolog antibody NGL antibody p185erbB2 antibody Proto-oncogene c-ErbB-2 antibody Proto-oncogene Neu antibody Receptor tyrosine-protein kinase erbB-2 antibody TKR1 antibody Tyrosine kinase type cell surface receptor HER2 antibody Tyrosine kinase-type cell surface receptor HER2 antibody V erb b2 avian erythroblastic leukemia viral oncogene homolog 2 (neuro/glioblastoma derived oncogene homolog) antibody V erb b2 avian erythroblastic leukemia viral oncogene homolog 2 antibody V erb b2 avian erythroblastic leukemia viral oncoprotein 2 antibody V erb b2 erythroblastic leukemia viral oncogene homolog 2 neuro/glioblastoma derived oncogene homolog (avian) antibody V erb b2 erythroblastic leukemia viral oncogene homolog 2 neuro/glioblastoma derived oncogene homolog antibody Verb b2 erythroblastic leukemia viral oncogene homolog 2 neuro/glioblastoma derived oncogene homolog (avian) antibody |
| Molecular Weight(MW) | 138kDa(Observed band size:180kDa) |
| Cellular Localization | Cell membrane, Cell projection, ruffle membrane; Early endosome, Cytoplasm, perinuclear region, Nucleus. |
WB
Western blot analysis of Phospho-HER2 / ErbB2 (Y1139) on different lysates with Rabbit anti-Phospho-HER2 / ErbB2 (Y1139) antibody at 1/1000 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 λpp 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 µg/Lane. Exposure time: 10 seconds; 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 at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of A431 cells treated with or without 100ng/mL EGF for 10 minutes labeling Phospho-HER2 / ErbB2 (Y1139) with Rabbit anti-Phospho-HER2 / ErbB2 (Y1139) antibody at 1/5000 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-Phospho-HER2 / ErbB2 (Y1139) antibody at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (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 (594) was used as the secondary antibody at 1/1000 dilution.
FC
Flow cytometric analysis of A431 cells (left) and A431 cells treated with 100ng/mL EGF for 10 minutes (right) labeling Phospho-HER2 / ErbB2 (Y1139). Cells were fixed and permeabilized. Then stained with the primary antibody (1/10000) (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 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000 ICC/IF:1:5000 FC:1:10000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
