Phosphotyrosine (0) antibody

Antigen: Phosphotyrosine (0)

Clonality: Monoclonal (PY20)

Host: Mouse

Reactivity: All Species

Application: Flow Cytometry (FACS), Immunofluorescence (IF), Immunoprecipitation (IP), Western Blotting (WB)

Catalog no.: ABIN179755

Additional Information

Alternative name: Phosphotyrosine

Immunogen: KLH-conjugated phosphotyrosine

Format: Purified

Isotype: IgG2b, kappa chain  (Matching secondary antibodies)

Clone: PY20

Description: Phosphorylation is a common modification of proteins that can result in alterations in protein function, protein-protein association, cellular localization, and protein-half life. Phosphorylation can occur on threonine, serine, and tyrosine residues. The PY20 monoclonal antibody recognizes phosphorylated tyrosine residues in all species tested (human, mouse, rat, dog, chicken, and frog). The PY20 antibody has been shown to be useful for flow cytometry, immunoprecipitation, Western blotting, and immunofluorescence staining. Hela cell extract was resolved by electrophoresis, transferred to nitrocellulose, and probed with monoclonal anti-phosphotyrosine antibody (clone PY-20). Lane 1, serum-starved Hela cells, Lane 2, serum-starved Hela cells following serum addition for 4 hrs. Lane 2 shows an upregulation of tyrosine phosphorylated proteins after serum addition. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system. Distribution: Phospho-Specific Function: Phosphorylation of specific tyrosine residues, signal transduction, cell cycle progression, oncogenic transformation

Application Details

Application Notes: Additional reported applications (for the relevant formats) include: immunoprecipitation, Western blotting, immunofluorescence microscopy. Western blotting, suggested working dilution(s): Use 1-2 µg per ml antibody dilution buffer per mini-gel. Do not use dilution or blocking buffers containing milk as they may interfere with antibody binding to proteins of interest. Dilution and blocking buffers containing 4% bovine serum albumin are recommended for use with this antibody. It is recommended that the reagent be titrated for optimal performance for each application.

Purification: The antibody was purified by affinity chromatography.

Buffer: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 50% glycerol. Final antibody concentration is 0.5 mg/ml.

Storage: The antibody solution should be stored at -20°C.

Research Area: Amino Acids, Protein Modifications

Restrictions: For Research Use only

Images

Hela cell extract was resolved by electrophoresis, transferred to nitrocellulose, and probed with monoclonal anti-phosphotyrosine antibody (clone PY-20). Lane 1, serum-starved Hela cells, Lane 2, serum-starved Hela cells following serum addition for 4 hrs. Lane 2 shows an upregulation of tyrosine phosphorylated proteins after serum addition. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system.

Publications

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

Vuori, Ruoslahti: "Tyrosine phosphorylation of p130Cas and cortactin accompanies integrin-mediated cell adhesion to extracellular matrix." in: The Journal of biological chemistry, Vol. 270, Issue 38, pp. 22259-62, 1995 (PubMed).

Prahalad, Calvo, Waechter et al.: "Regulation of MDCK cell-substratum adhesion by RhoA and myosin light chain kinase after ATP depletion." in: American journal of physiology. Cell physiology, Vol. 286, Issue 3, pp. C693-707, 2004 (PubMed).