		Technical Data
E3451-35	ErbB3 (ERBB3, v-erb-b2 Erythroblastic Leukemia Viral Oncogene Homolog 3 avian, HGNC:3431, ErbB-3, HER3, MDA-BF-1, c-erbB-3, c-erbB3, erbB3-S, p180-ErbB3, p45-sErbB3)

Description:
c-erbB-3/HER-3 is a member of the EGF receptor tyrosine kinase family. It is a kinase defective receptor which signals by forming heterodimers with other erb family members. c-erbB-3 is over-expressed in a variety of tumors including breast, stomach, pancreas and colon, although the significance is unclear because over-expression of c-erbB-3 has no effect on tumorigenic growth of cells. Heregulin and EGF stimulate tyrosine phosphorylation of c-erbB-3 to different extents. This antibody can be used to assess the contribution of the other erb family members co-expressed with c-erbB-3 to ligand-dependent signaling because it inhibits the binding of heregulin (NDF) to c-erbB-3/HER-3.

Applications:
Suitable for use in Immunocytochemistry, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.

Recommended Dilutions:
Immunocytochemistry: 10ug/ml shows positive immunostaining for erbB-3 in MCF-7 cells fixed with 95% ethanol/5% acetic acid.
Western Blot: 0.1-2ug/ml detects erbB-3 (~185kD) in RIPA lysates of human MCF-7 cells; previous lots have detected erbB-3 in MDA RIPA lysates. MCF-7 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with E3451-35. Proteins were visualized using an IgG (HRP) goat anti-mouse and a chemiluminescence detection system.
Immunoprecipitation: 4ug antibody immunoprecipitates erbB-3 from human MDA and MCF-7 RIPA lysates.
Immunohistochemistry: 1:50-1:100 dilution of a previous lot detected erbB-3 membrane staining in human normal and carcinoma breast tissue.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Type	Isotype	Clone	Grade
Mab	IgG2a,k	0.T.39	Affinity Purified

Size		Storage	Shipping	Source	Host
200ug		-20°C	Blue Ice	Human	Mouse

Concentration:

~0.2mg/ml

Immunogen:

Recombinant rat erbB-3/HER-3 oncoprotein expressed in insect cells and purified by metal chelating chromatography. Epitope maps between aa1295-1323 of human erbB-3 (NM_001982).

Purity:

Purified by Protein A affinity chromatography.

Form

Supplied as liquid in 10mM PBS, pH 7.4, 0.2% BSA, 0.09% sodium azide.

Specificity:

Recognizes human erbB-3 (~185kD). Does not crossreact with erbB-1 (EGFR) and erbB-2/HER-2. Species Sequence Homology: mouse, rat and bovine.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

1. Keely, S.J. & Barrett, K.E., J. Mol. Biol. 274: 33,449-33,454 (1999). 2. Ezeh, P.I. & Farbman, A.I., J. Neurobiol. 37: 199-210 (1998). 3. Kraus, M.H., Proc. Natl. Acad. Sci. USA 86: 9193-9197 (1989). 4. Paolo, F., Mol. Cell. Bio. 14: 492–500 (1994).