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09-18-2007, 09:18 AM
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 washing of phosphorylated plasma protein All proteins in a plasma sample is precipitated with 35% ACETONE/WATER and centrifuged. Then I want to wash the protein pellet one or several times to make sure that there is no free phosphate (from the plasma) associated with the pellet. After this I dissolve the pellet in 1 M NaOH and heat to 70C to release all (alkali labile) protein bound phosphate for quantification. My question: What buffer should I wash my protein pellet with that ensures complete removal of free phosphate and plasma but at the same time without releasing serine-bound phosphate? Many thanks to you who can help a PhD student in analytical chemistry with little experience of working with blood and proteins.        |
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