I'm working on subcellular fractionation. I use Vinculin, Lamin B1, and COX IV antibody for cytoplasmic, nuclear, and mitochondrial fraction, respectively. However, I'm not quite sure if I have plasma membrane contamination in my mitochondrial lysates. Therefore, I was wondering if anyone could suggest a control antibody for the protein which is present in plasma membrane but not in mitochondrial membrane?
Thank you very much
you could try an antibody against any protein known to be on the cell surface but not the mitochondria. One of many would be a caveolin.
I agree with Bruce Allen, but you should also include a marker protein for contamination of the mitochondrial fraction by endo- or sarcoplasmic reticulum. Mitochondria are often located near the endo- or sarcoplamic reticulum and interact with them. The suggested antibodies depend on your organism but SERCA ATPase or calreticulin could work well.
Good luck!
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