Phosphotyrosine antibody (Biotin)

Antigen: Phosphotyrosine

Clonality: Monoclonal (PY20)

Host: Mouse

Reactivity: Please enquire

Conjugate: Biotin

Application: Immunofluorescence (IF), ELISA, Immunoprecipitation (IP), Western Blotting (WB)

Catalog no.: ABIN135061

Additional Information

Isotype: IgG2b

Clone: PY20

Description: Phosphorylated tyrosine and phenylphosphate Protein tyrosine residues are phosphorylated as a result of intracellular protein kinase activation (e.g. via growth factors) during normal growth and development, and in oncogenesis. The most abundant population of target proteins for tyrosine phosphorylation are cell surface glycoproteins. Antibody to phosphotyrosine enables the detection, isolation and characterization of proteins containing phosphotyrosine. PY20 also prevents internalization of activated receptors, e.g. EGFr, when microinjected into cells. The affinity of PY20 for phosphotyrosine is approximately 10-6 to 10-7 M. PY20 binding to phosphorylated tyrosines can be inhibited by free phosphotyrosine and phenylphosphate, but not by phosphoserine, phosphothreonine or free phosphate.

Application Details

Application Notes: WORKING DILUTIONS Western Blotting:Purified ( 2 ug/mL) ELISA:Purified (100 nug/mL), AP (1:500-1:1,000); HRP (1:1,000-1:4,000), Immunoaffinity purification: Purified antibody 4 mug/mL Sepharose-4B. Since applications vary, you should determine the optimum working dilution for the product that is appropriate for your specific need.

Purification: Supernatant

Storage: The purified (UNLB) antibody is supplied as 0.5 mg of purified immunoglobulin in 1.0 mL of 100 mM borate buffered saline, pH 8.0. No preservatives or amine-containing buffer salts added. Store at 2-8C. The fluorescein (FITC) conjugate is supplied as 0.5 mg in 1.0 mL of PBS/NaN3. Store at 2-8C. The alkaline phosphatase (AP) conjugate is supplied as 1.0 mL of stock solution in 50mM Tris/1mM MgCl2/50% Glycerol, pH 8.0, containing 0.1% NaN3 as preservative. Store at 2-8C or long-term at -20C. The horseradish peroxidase (HRP) conjugate is supplied as 1.0 mL of stock solution in 50% glycerol/50% PBS, pH 7.4. No preservative added. Store at 2-8C or long-term at -20C. The biotin (BIOT) conjugate is supplied as 0.5 mg in 1.0 mL PBS/NaN3. Store at 2-8C. The R-phycoerythrin (R-PE) conjugate is supplied as 0.1 mg in 1.0 mL of PBS/NaN3 and a stabilizing agent. Store at 2-8C. Do not freeze! Reagents are stable for the period shown on the label if stored as directed. WARNING Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.

Research Area: Signaling, Metabolism, Amino Acids, Protein Modifications

Restrictions: For Research Use only

Publications

Takagi, Daibata, Last et al.: "Intracellular localization of tyrosine kinase substrates beneath crosslinked surface immunoglobulins in B cells." in: The Journal of experimental medicine, Vol. 174, Issue 2, pp. 381-8, 1991 (PubMed).

Glenney, Zokas, Kamps: "Monoclonal antibodies to phosphotyrosine." in: Journal of immunological methods, Vol. 109, Issue 2, pp. 277-85, 1988 (PubMed).