Prove for subcellular fractionation?

Thorsten Schmidt Thorsten.Schmidt at rz.ruhr-uni-bochum.de
Sun May 18 11:10:28 EST 1997


Dear reader!

I made a fractionated protein isolation of cells and got
fractions that are told to be 
1. "nuclear"
2. "mitochondrial/synaptosomal"
3. "microsomal", 
4. "cytosolic",
But how can I prove that they really contain that and that perhaps
no other fractions contains mitochondrial proteins?

I´ve been told that one uses
citrate synthase, lactate dehydrogenase and choline phosphotransferase
as "marker enzymes" but how?
Can you tell me special assays for them?
There are no antibodies availible which detect them!

Do you know special proteins which should be only present in one of
these fractions and can be detected by commercially availible antibodies?

Do you perhaps have a good and working protocol for fractionated
protein isolation?

It would be too kind if you´ll send me an answer.

Thank you so much in advance

Thorsten Schmidt




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