Anti-erbB-3/HER-3 Antibody, clone 2F12

185 kDa

The ErbB3 gene encodes a member of the epidermal growth factor receptor (EGFR) family of receptor tyrosine kinases. ErbB3 is a membrane-bound protein which has a neuregulin binding domain but not an active kinase domain. It can therefore bind this ligand but cannot convey a signal into the cell via protein phosphorylation. However it does form heterodimers with other EGF receptor family members which do have kinase activity. Heterodimerization leads to the activation of pathways which lead to cell proliferation or differentiation. Amplification of this gene and/or overexpression of its protein have been reported in numerous cancers including prostate, bladder and breast tumors. Alternate transcriptional splice variants encoding different isoforms have been characterized. One isoform lacks the intermembrane region and is secreted outside the cell. This form acts to modulate the activity of the membrane-bound form. Additional splice variants have also been reported but they have not been thoroughly characterized.

Epitope: a.a. 1295-1323 of human erbB-3

Recombinant rat erbB-3/HER-3 oncoprotein expressed in insect cells and purified by metal chelating chromatography. The epitope maps between amino acid residues 1295-1323 of human erbB-3. Clone 2F12.

Detect erbB-3/HER-3 using this Anti-erbB-3/HER-3 Antibody, clone 2F12 validated for use in IC, IH, IP & WB.

Immunoprecipitation:
4 µg of a previous lot of this antibody immunoprecipitated erbB-3/HER-3 from human MDA and MCF-7 RIPA lysates.

Immunocytochemistry:
10 µg/mL of a previous lot of this antibody gave positive immunostaining for erbB-3/HER-3 in MCF-7cells fixed with 95% ethanol/5% acetic acid.

Immunohistochemistry:
1:50 and 1:100 dilution of a previous lot detected erbB-3/HER-3 membrane staining in human normal and carcinoma breast tissue.

Research Category
Signaling

Research Sub Category
Growth Factors & Receptors

Predicted to cross-react with mouse, rat, and cow.

This antibody recognizes erbB-3/HER-3, Mr 185 kDa. Does not cross-react with erbB-1 (EGFR) and erbB-2/HER-2.

Format: Purified

Protein A chromatography

Purified mouse monoclonal IgG2aκ in buffer containing 10 mM PBS, pH 7.4, 0.2% BSA, with 0.09% sodium azide. Frozen solution.

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Control
Human breast adenocarcinoma (well-differentiated), mouse brain lysate, MCF-7 cell lysate.

Routinely evaluated by Western Blot on RIPA lysates of human MCF-7 cells.

Western Blot Analysis:
0.1-2 µg/mL of this lot detected erbB-3/HER-3 in RIPA lysates of human MCF-7 cells. Previous lots have detected erbB-3/HER-3 in MDA RIPA lysates.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

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