Application
Extracts of SK-BR-3 cells were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was either not treated (1-5, 7) or treated (6, 8) with YOP phosphatase, blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with the ErbB-2 [pY1248] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5-8), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphotyrosine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab)2 anti-rabbit IgG alkaline phosphatase and signals were detected.The data show that only the phosphopeptide corresponding to ErbB-2 [pY1248] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, further verifying that the antibody is phospho-specific.| Product Name | Her2 / ErbB2 (phospho Tyr1248) antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal antibody to ErbB2 (phospho Tyr1248)ErbB2 (also known as Her2 or neu) is a 185 kDa transmembrane receptor tyrosine kinase from the EGFR family that acts to regulate a variety of biological responses including cell growth, differentiation, and tissue development. Ligand binding to the extracellular domain, or overexpression of the receptor leads to phosphorylation on several tyrosine residues within the cytoplasmic domain, and activation. Overexpression or abnormal activation of ErbB2 has been found in a variety of tumors including brain, breast, lung and skin cancer. Tyrosine 1248 within the cytoplasmic domain of the receptor is an auto-phosphorylation site that allows binding of Shc and activation of the Ras, Raf, ERK1&2 signaling pathway. |
| Immunogen | Synthetic phosphopeptide derived from a region of human ErbB-2 that contains tyrosine 1248. |
| Modification | p-Tyr1248 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | WB |
| WB: Use at a concentration of 0.1 - 1.0 μg/ml. Detects a band of approximately 185 kDa. Optimal dilutions/concentrations should be determined by the end user.: |
|
| Species Reactivity | HumanDogRatHamster |
| Concentration | 0.5mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | c-erb B2 Herstatin HER2 137215 cerb B2 ErbB2 NGL HER2/neu P04626 2064 NEU HER-2/neu HER-2 TKR1 HER 2 c erb B2 HER 2/neu ErbB2 (phospho Y1248) ErbB2 (phospho Tyr1248) ErbB2 (pY1248) ErbB2 (pTyr1248) ErbB2 phospho Y1248 ErbB2 phospho Ty |
|---|---|
| Target | ErbB2 |
| Cellular Localization | Cell Membrane |
Application
Extracts of SK-BR-3 cells were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was either not treated (1-5, 7) or treated (6, 8) with YOP phosphatase, blocked with a 5% BSA-TBST buffer overnight at 4°C, then incubated with the ErbB-2 [pY1248] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5-8), the non-phosphopeptide corresponding to the phosphopeptide immunogen (2), a generic phosphotyrosine-containing peptide (3), or the phosphopeptide immunogen (4). After washing, the membrane was incubated with goat F(ab)2 anti-rabbit IgG alkaline phosphatase and signals were detected.The data show that only the phosphopeptide corresponding to ErbB-2 [pY1248] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, further verifying that the antibody is phospho-specific.| Positive Control | SK-BR-3 cell line (overexpresses ErbB-2) |
|---|---|
| Application Notes | WB: Use at a concentration of 0.1 - 1.0 μg/ml. Detects a band of approximately 185 kDa. Optimal dilutions/concentrations should be determined by the end user.: |
| Form | Liquid |
|---|---|
| Storage Instructions | Keep as concentrated solution. Aliquot and store at -20ºC or below. Avoid freeze-thaw cycles. |
| Storage Buffer | Preservative: 0.05% Sodium Azide; Constituents: PBS, 1mg/ml BSA. pH 7.3 |
