HER3 antibody

• Catalog Number: orb344608
• Category: Antibodies
• Description: HER3 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IP, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: 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.
• Concentration: 2.5 mg/mL
• Dilution range: ELISA: 1:10,000 - 1:50,000, IP: Use 5 µL antibody., WB: 1:500- 1:2,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: 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.
• Conjugation: Unconjugated
• UniProt ID: P21860
• NCBI: NP_001005915.1
• Storage: 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.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-HER3 Antibody, HER, c erbB 3 antibody,
• Note: For research use only
• Application notes: 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.
• Expiration Date: 12 months from date of receipt.

Combined immunoprecipitation and western blot of a human cell lysate using anti-HER3 antibody. Lysates were prepared from a human LNCap prostate cell line both with (+) treatment with Heregulin-b1 for 15' at 100 ng/mL and without (-) the addition of Heregulin-b1. The combination of immunoprecipitation and western blotting was performing using 5 µL of the anti-HER3 antibody for immunoprecipitation followed by western blot detection using an anti-phosphotyrosine antibody. Visualization occurred using an ECL system. Film exposure approximately 1'. Other detection systems will yield similar results.

Signaling pathways activated by the ErB3/HER3 Receptor.