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Western blot analysis was performed on whole cell extracts (30 µg lysate) of Jurkat (Lane 1), Jurkat treated with Sodium vanadate (1mM for 30 minutes) (Lane 2), Raji (Lane 3) and Raji treated with Sodium vanadate (1mM for 30 minutes) (Lane 4). The blots were probed with Anti-Phosphotyrosine / pY Rabbit Polyclonal Antibody, HRP conjugate (Product # 61-5820, 2 µg/ml). Bands corresponding to phosphorylated tyrosines were observed only upon treatment with Sodium vanadate in the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Chemical|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Rabbit anti-Phosphotyrosine-HRP; HRP conjugated Polyclonal antibody specific to Multiple Phosphotyrosine [pY]. This antibody is validated for use in Western Blot, Immunoassay (ELISA).|
|Storage buffer||PBS, pH 7.4, with 4mg/ml BSA, 40% glycerol|
|Contains||0.1% Proclin 300|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
The role of tyrosine phosphorylation in transduction of the mitogenic signal from transmembrane receptors and in transformation by oncogene tyrosine kinases has been the subject of intense investigation for several years. While the phosphorylation of specific tyrosine residues has been shown to be a primary mechanism of signal transduction during normal mitogenesis, cell cycle progression and oncogenic transformation, its role in other areas such as differentiation and gap junction communication, is a matter of active and ongoing research. Antibodies that specifically recognize phosphorylated tyrosine residues have proved to be invaluable to the study of tyrosine -phosphorylated proteins and the biochemical pathways in which they function. The fluorescein (FITC) conjugate of clone PY20 anti-phosphotyrosine is especially useful for the detection of these P-Tyr proteins in immunohistochemical and immunocytochemical protocols in situations wherein the use of a secondary antibody would complicate detection of the protein(s) of interest.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Human||Not Cited||The role of signal transducer and activator of transcription 5 in the inhibitory effects of GH on adipocyte differentiation.||Richter HE,Albrektsen T,Billestrup N||Journal of molecular endocrinology (30:139)||2003|
Phosphotyrosine; pTyr; pY