How to concentrate the protein
Posted 05 July 2010 - 10:49 PM
I need huge concentration of my protein for my assays. For this I lysed transiently transfected 293T cells with 1%SDS and the lysate was further diluted with RIPA buffer containing 1X PBS. the lysate was lyophilized. lyophilized product was dissolved in RIPA buffer. But this protocol is creating problem, becoz of high salt concentration SDS PAGE gel run is not proper(there is a drag,which brings smear). there is not much difference in protein concentration of input and lyophilized sample.
can someone suggest how to concentrate as well as get rid of salts from the lysate. Bcoz at the end of the day I need to send the samples for mass spec.
Posted 07 July 2010 - 07:25 AM
Posted 08 July 2010 - 03:28 AM