Purifying nuclear fraction from skeletal muscle homogenate
Posted 04 May 2009 - 11:50 AM
Our current protol:
We homogenize each sample on high in a buffer that also contains 1% Tritonx100. We let the samples sit on ice for 1 hour and then centrifuge at 13,000G for 30mins. The supernatant is collected and labled as the cytosolic and membrane bound protein fraction which is used for western blotting. I am assuming the the pellet contains myofibrillar, connective tissue, and possibly mitchondria and nuclear fraction (?).
My first question is if my assumptions are correct on what is found in each fraction? Second, is there a protocol to purify the nuclear proteins from the pellet?
Posted 07 May 2009 - 08:53 AM
Understanding Life One protein at a time...
The Peptide company
Understanding Life One Protein At a Time...