HER3 Antibody
Anti-HER3 Antibody - Immunoprecipitation/Western Blot
Signaling pathways activated by the ErB3/HER3 Receptor.
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Datasheet

HER3 Antibody

Rabbit Polyclonal

200-401-205
250 µg
ELISA, IP, WB
Human
Rabbit
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Description

Background

HER3 antibody is ideal for western blotting, ELISA and IP. The human Epidermal Growth Factor Receptor (EGFR) family is a family of receptor tyrosine kinases consisting of 4 members: EGFR, HER2, HER3 and HER4.  These receptors are expressed on a wide variety of tissue types and have been implicated in many human cancers, including breast, gastric, colon and prostate.  Strong expression of both HER3 and HER2 (also known as p185neu) by tumor cells can be associated with poor outcome for patients.  The ligand for HER3 and HER4 is called Neu Differentiation Factor (NDF) or Heregulin.  In some instances, antibodies against these receptors prove effective in combination therapy regimens for treatment of the disease.  Signaling through the EGFR family involves the combinatorial dimerization of the four known family members and is mediated by at least 8 different hormones, including EGF and the heregulins.  Heregulin binding and tyrosine phosphorylation by HER3 or HER4 is enhanced in the presence of HER2.  Structurally the members of the EGFR family are 1210 to 1342 amino acids in length with approximately 650 amino acids of the extracellular domain being heavily glycosylated.  The extracellular domains show up to 70% sequence homology.

Application Note

Anti-HER3 antibody has been tested by ELISA, immunoprecipitation, and western blot and is specifically designed for immunoprecipitation and can be used for immunoblotting in conjunction with a detection antibody like anti-phosphotyrosine.  Reactivity in other assays is likely, but has not been determined.  Recognition of HER3 is independent of the phosphorylation status of the molecule.  Human prostate cell lines (LNCap) or human breast cell line (MDA-453) are typically used as positive control sources. For immunoprecipitation use approximately 5 µL of the antibody. The immunoprecipitation mix contained the antibody, 25 µL of Protein A-agarose beads and 1.0 ml of lysate (lysate contains approximately 1.0 mg of total protein). This mixture is rotated overnight at 4°C and then washed 3 times with lysis buffer (used to prepare the lysate). The resulting bead complex is dissolved in 20-30 µL of 3X SDS-PAGE sample buffer and approximately 15 µL is loaded per lane on an 8% polyacrylamide gel. The combination of immunoprecipitation and immunoblotting was performing using the anti-HER3 antibody for immunoprecipitation followed by immunoblot detection using an anti-phosphotyrosine antibody. The complex is then reacted with HRP Goat-anti-Rabbit IgG and ECL for detection and shows a single HER3 band at 180kDa. Researchers should determine optimal titers for other applications.

Purity/Specificity

Rabbit Anti-HER3 is an IgG fraction antibody purified from monospecific antiserum by Protein A chromatography followed by cross adsorption against GST and extensive dialysis against the buffer stated above.  Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.  The antibody is directed against human HER3 and is useful in determining its presence in immunoprecipitation experiments. This antibody can detect HER3 from human, mouse and rat sources.   Reactivity of this antibody with HER3 from other species is unknown.

Anti-HER3 (RABBIT) Antibody - 200-401-205
rabbit anti-HER3 Antibody, HER, c erbB 3 antibody, c erbB3 antibody, ERBB3 antibody, ERBB3 protein antibody, erbB3 S antibody, Glial growth factor receptor antibody
ERBB3
ELISA, IP, WB
Human
Human
Rabbit
Polyclonal
IgG
Recombinant Protein
Anti-HER3 whole rabbit serum was prepared by repeated immunizations with a HER3 fusion protein corresponding to amino acids 1283 to 1323 (40 amino acids) at the carboxy-terminus of human HER3.
Liquid (sterile filtered)
2.5 mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
None
Dry Ice
Store Anti-HER3 at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Expiration date is one (1) year from date of opening.

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This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.

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