Western blot does not match IF
Posted 03 April 2012 - 12:42 PM
I have seen two posts on here with this problem, but the answers have not helped so far...
I get nuclear protein (about 90%) with immunofluorescence, but using the same antibody I get 95% cytoplasmic protein in western blot (NE-PER kit). My extractions are very clean, i have used antibodies to measure this.
The only thing I can think of is that SIRT1 my molecule of interest is somehow leaving the nucleus during the extraction.
I am going to try cross-linking with formaldehyde, and glycine.
Can anyone shed light on this?
any ideas would be much appreciated
Posted 03 April 2012 - 05:12 PM
Posted 03 April 2012 - 10:31 PM
I'm going to try crosslinking before extraction, has anyone used this?