problems with detecting protein expression - protein expression in cell (Mar/18/2009 )
I am having my research on transcription of microorganism.
in order to detect the protein expression in cell, either in cytoplasm or in the nucleus,
anyone could help me this.
i have already find out the method by using flurosence marker and observe with confocol,
but i have no confocol microscopy in lab, any other "classical" method to proof the protein either express in cytoplasm or in nucleus?
Micro-organisms don't have a nucleus... and organisms that do have a nucleus don't have ribosomes in their nucleus. So are you interested in the final location of the proteins, rather than where they are expressed?
swanny on Mar 18 2009, 03:01 PM said:
i am doing virus replication, and try to find out where the protein are in the cell by using cell culture.
You might be able to get something by doing a gentle lysis, then separating the nucleus from the cytoplasm, and running gels to look for your protein(s) in the two.
I think Swanny's method of differential centrifugation will work. This website may work http://faculty.plattsburgh.edu/donald.slish/DiffCent.html . Good Luck