When I perform HeLa cell nucleus-cytoplasm seperation, I use human MSH2 as a marker protein for nucleus fraction. But the western blot show a sharp strong band in cytoplasm and nothing in nucleus. The antibody is bought from abcam (ab92473). Other marker proteins are all normal as predicted (HSP90 for cyto, HDAC1 for nuc). Could the antibody be mislabled? but the molecular weigh is correct.
for nuclear proteins translocation to cytoplasm is a method to regulate activity. Further more you should recheck you protein fractionation protocol
regards
Sven
Small amounts of MSH2 can be found at the cytoplasm. But most of the MSH2 is located at the nucleus. Santa Cruz and BD have good antibodies for MSH2 is you doubt about the antibody. However, in my opinion you should recheck your protocol it seems that your fractionation is not correct.
Janica and Sven, thanks for your help. But like I said, MSH2 is not my only marker protein, I also checked with HSP90 MEK1 (for cyto) and HDAC1 HDAC2 H2B (for nuc). And all these marker performed just well. So I think there is nothing wrong with my protocol.
Well then is very strange If is not your protocol. Because I have been doing nuclear extract for in vitro MMR and I never lost MSH2 into the cytosollic fraction.
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