Open Forum on Methodology
12 July 2005
July 11, 2005
Question on cell fractionation:
Does anyone have a good protocol (or suggestion, or explanation) for cell fractionation in order to separate nuclear from cytoplasmic fractions? We are trying to show that a protein that shows a clear nuclear localization in immunofluorescence assay can be found predominantly in the nuclear fraction. However, using the cell fractionation protocol we have, the protein shows equal distribution between cytoplasm and nucleus, whereas the control proteins (tubulin and PARP) show the expected distribution. We think that probably our protein is loosely present in the nucleus and becomes easily leaky during the process.
Thank you for your help and suggestions.
Protocol being used:
Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882