Phosphotyrosine

Phosphorylation and dephosphorylation of cellular proteins are central steps in transducing extracellular signals to the cell nucleus. Phosphorylated epitopes may serve as docking sites for the assembly of protein complexes or may alter the 3-dimensional protein structure thus modulating enzymatic activity or the ability to undergo protein-protein interactions. Modification of proteins on tyrosine residues is mediated by protein tyrosine kinases. \n\nApplications: \nSuitable for use in ELISA, Immunoprecipitation, and Western Blot. Other applications not tested.\n\nRecommended Dilution:\nELISA: 0.05ug/ml\nWestern Blot: 1ug/ml for AP/BCIP/NBT (MTT) detection; 0.1ug/ml for HRP/ECL detection\nImmunoprecipitation: 1-10ug per 10e6 vanadate-treated A431 cells \nOptimal dilutions to be determined by the researcher.\n\nPositive Control (supplied): \nPhosphotyrosine Molecular Weight Marker. Supplied as a lyophilized powder. Reconstitute with 200ul sterile, ddH2O. After complete solubilization of the proteins, add 200ul SDS-PAGE sample buffer and incubate at 90 degrees C for 5 minutes. For WB applications, use 20ul/lane (mini gel) for HRP/DAB detection and 10ul/lane (mini gel) for HRP/ECL detection. Do not use NC membranes.\n\nStorage and Stability:\nLyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile ddH2O. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

P4078-05E

Size

100ug

Applications

ELISA, IP, WB

Hosts

Mouse

Form

Supplied as a powder lyophilized from PBS containing PEG, sucrose, 0.09% sodium azide. Reconstitute with 1ml ddH2O.

P Type

Mab

Purity

Purified by thiophilic adsorption and size exclusion chromatography

Isotype

IgG1,k

Additional Info

Recognizes a broad spectrum of tyrosine-phosphorylated proteins in crude cell extracts and may be particularly well suited for the detection/screening of tyrosine phosphorylated proteins.