Jurkat cells are treated with 100 nM calyculin A and 1 mM pervanadate to inhibit multiple serine/threonine and tyrosine phosphatases, respectively and upregulate protein phosphorylation. Treated Jurkat cells were lysed in 1X cell lysis buffer and lysates were lyophilized.
Applications
The recommended final concentration for this positive control lysate is 1.0 mg/ml. Reconstitute the lyophilized positive control with 225 μl dH2O to create a 6.0 mg/ml lysate solution. Dilute the sample with 1.0 ml 1X cell lysis buffer (#9803) or a sample diluent supplied with one of our PathScan® Sandwich ELISA kits, to make the final concentration at 1.0 mg/ml. Keep on ice. 100 μL samples should be used in each well of a 96-well plate.
Background
PathScan® Sandwich ELISA is a convenient tool for the analysis of protein expression as well as post-translational modifications (such as phosphorylation and acetylation) allowing the researcher to study signal transduction. PathScan® Sandwich ELISA Control Phospho Cell Extracts II can be used not only to troubleshoot the experiment but also provides the means to standardize the signal allowing the comparison of results obtained from different plates or experiments.
注意事项:
Storage: Supplied as lyophilized product. Keep at -20ºC for
short term storage or at -80ºC for longer periods. Avoid repeated
freeze-thaw cycles