membrane proteomics - (Apr/07/2013 )
I have started working on membrane proteomics and have planned to do BNPAGE according to Schagger et al (2006). I have a problem that we dont have a centrifuge going upto 100,000xg. can we separate the cell membrane proteins by increasing the time of centrifugation decresing the speed. Secondly is any one is doing BNPAGE i need some guidance regarding that too.
theoretically, you can reduce rcf and increase time to get the same result. you can calculate the time necessary if you know the sedimentation coefficient of the material you wish to pellet or you can use "trial and error" to determine the necessary time.
bear in mind that most centrifugation reported in the literature is excessive (to ensure completion).