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I ordered PKC alpha protein (Active).(ab60839, Serial no: k1111-900) from your company, and want to use it to phosphorylate an E.Coli purified protein, but I could not find the protocol or the recipe of kinase reaction buffer for this kinase on your website. I also found there are different recipe of kinase reaction buffer for PKC alpha protein on the web. So could you suggest a standard protocol or a recipe of kinase reaction buffer for this kinase for phosphorylating a E.Coli purified protein?
Asked on Jun 05 2012
Thank you for contacting us.
Please find attached a modified version of the radiolabel (phosphorus 33-ATP) protocol that was used to measure activity of ab60839. The assay utilized CREBtide as the substrate (KRREILSRRPSYR) but the lab believes you can simply substitute your E. coli protein for the peptide, which I have noted in bold at the bottom of the materials section of the protocol.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Answered on Jun 05 2012