Standard Antibodies Against Separated Cellular Preps - (Aug/25/2006 )
I am new to immunology and was wondering if anyone could tell me if there is a standard antibody used when checking you have a 1) clean nuclear prep 2) clean cytosolic prep and 3) clean membrane prep. I have separated my cellular prep into these three groups but want to check they are clean on a Western.
i don't regulary deal with membrane preps, but you can use a protein for cytosolic protein or nuclear protein.
For ex, TBP is a good nuclear specific protein.
"clean" is too ambitious, what you can do is determing the enrichment factor of each compartment; use of reference proteins dependes on cell type, f.i. a standard against epthelial basal membrane is Na-K-ATPase, for apical IQGAP will work; for cytosol use a glycolytic protein, DNA is a good reference for the nucleus; try to find out was is commonly used for your special cell or tissue type