Human Phospho-ErbB2/Her2 (Y1196) Antibody

Catalog #: AF4438 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF4438
AF4438-SP

ErbB2/Her2 in Human Breast Cancer Tissue.

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Human Phospho-ErbB2/Her2 (Y1196) Antibody Summary

Species Reactivity

Human

Specificity

Detects human ErbB2/Her2 when phosphorylated at Y1196 in Western blots.

Source

Polyclonal Rabbit IgG

Purification

Antigen Affinity-purified

Immunogen

Phosphopeptide containing human ErbB2/Her2 Y1196 site

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

See below

Simple Western

5 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry

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ErbB2/Her2 in Human Breast Cancer Tissue. ErbB2/Her2 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human Phospho-ErbB2/Her2 (Y1196) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4438) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western Blot

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Detection of Human Phospho-ErbB2/Her2 (Y1196) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with the indicated concentration of Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 0.1 µg/mL Rabbit Anti-Human Phospho-ErbB2/Her2 (Y1196) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4438) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-ErbB2/Her2 (Y1196) was detected at approximately 185 kDa (as indicated). For additional reference, duplicate lysates were probed with 1 µg/mL Human ErbB2/Her2 Monoclonal Antibody (lower panel, Catalog # MAB1129). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western

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Detection of Human Phospho-ErbB2/Her2 (Y1196) by Simple Western^TM. Simple Western lane view shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 10 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-ErbB2/Her2 (Y1196) at approximately 237 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human Phospho-ErbB2/Her2 (Y1196) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4438). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: ErbB2/Her2

ErbB2, also called Neu and Her2 (human epidermal growth factor receptor 2), is a type I membrane glycoprotein that is a member of the ErbB family of tyrosine kinase receptors. ErbB family members serve as receptors for the epidermal growth factor (EGF) family of growth factors. ErbB2 is widely expressed in epithelial cells and has also been found to be over-expressed in a large number of breast carcinomas. Among ErbB family members, ErbB2 is unique in that it has no identified ligands. Rather, ErbB2 heterodimerizes with the other members of the ErbB family (ErbB1 (EGF R), ErbB3, ErbB4) to form higher affinity signaling complexes. Because ErbB3 contains a defective kinase domain, the kinase domain of ErbB2 is responsible for initiating the tyrosine phosphorylation signal through the heterodimeric receptor. It has been found that a discrete three amino acid signal in the ErbB3 cytoplasmic domain is critical for transactivation of ErbB2. Interestingly, this same three amino acid signal has also been found in ErbB1 and ErbB4. Phosphoinositide 3-kinase has been shown to play a role in ErbB2 signal transduction. The cytoplasmic domain of ErbB2 has been shown to associate with beta-catenin and plakoglobin. Human ErbB2 consists of 1255 amino acids (aa) with a 21 aa signal sequence, a 631 aa extracellular domain, a 23 aa transmembrane region, and a 580 aa cytoplasmic domain. ErbB2 can be shed from the cell surface by proteolytic cleavage by an unidentified protease. ErbB2 appears to play roles in development, cancer, communication at the neuromuscular junction and regulation of cell growth and differentiation (1-10).

References

Coussens, L. et. al. (1985) Science 230:1132.
Yamamoto, T. et. al. (1986) Nature 319:230.
Kanai, Y. et. al. (1995) Biochem. Biophys. Res. Commun. 208:1067.
Codony-Servat, J. et. al. (1999) Cancer Res. 59:1196.
Carraway, K.L. 3^rd et. al. (1994) J. Biol. Chem. 269:14303.
Emkey, R. and C.R. Kahn (1997) J. Biol. Chem. 272:31172.
Schaefer, G. et. al. (1999) J. Biol. Chem. 274:859.
Schlessinger, J. (2000) Cell 103:211.
Hellyer, N.J. et. al. (2001) J. Biol. Chem. 276:42153.
Daly, R.J. (1999) Growth Factors 16:255.

Long Name

Receptor Tyrosine Protein Kinase ErbB2

Entrez Gene IDs

2064 (Human); 13866 (Mouse); 24337 (Rat); 102146608 (Cynomolgus Monkey)

Alternate Names

CD340 antigen; CD340; c-erb B2/neu protein; EC 2.7.10; EGFR2; ErbB2; HER2; HER-2; HER2EC 2.7.10.1; herstatin; Metastatic lymph node gene 19 protein; MLN 19; MLN19; Neu Oncogene; NEUHER-2/neu; neuroblastoma/glioblastoma derived oncogene homolog; NGL; NGLTKR1; p185erbB2; Proto-oncogene c-ErbB-2; Proto-oncogene Neu; receptor tyrosine-protein kinase erbB-2; TKR1; Tyrosine kinase-type cell surface receptor HER2; v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 2(neuro/glioblastoma derived oncogene homolog); v-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastomaderived oncogene homolog (avian)