question about protein molecular weight - (Dec/10/2007 )
I did native gel electrophoresis with 2-ME, and there appears to be have just one band, which is in the same position as the monomer on the native gel without 2-ME. What does it mean? Thank you.
Is it still necessary to do native gel with 2-ME? I think they are in the same mechanism.
You want to see if charged group attached to the thiol groups, right? SDS-PAGE will not show charge difference as sensitive as native gel.
Were the two bands well separated from each other in the native w/o 2-ME, or very close?
Now repeat an SDS-PAGE, do one without 2-ME on one side of the gel and one with 2-ME on the other side to minimize cross-over. Load the 2-ME sample at last after adding the MW markers, start the run without delay, both should have MW marker lanes nearby. If you see basically the same patern as the native gels with or without 2-ME, then I am more convinced that this is a dimerization, rather than charged group hocked on one of the thiols.
Please help me check this picture attached. Thank you!
Now repeat an SDS-PAGE, do one without 2-ME on one side of the gel and one with 2-ME on the other side to minimize cross-over. Load the 2-ME sample at last after adding the MW markers, start the run without delay, both should have MW marker lanes nearby. If you see basically the same patern as the native gels with or without 2-ME, then I am more convinced that this is a dimerization, rather than charged group hocked on one of the thiols.