Recently, when I do IF experiments, I find GFP tag without any target proteins always locates at centrosome though this kind of dot location is not so obvious, we still can see it. I fix the cells with cold-PFA on the ice for 10min then wash it with PBS 3 times for next step.
I did the experiments to fix GFP tagged target protein with cold methonal on the ice, but the signal is delocation. It should be a protein that locates on the cenrtrosme. However when I fix the cells with methonal I cannot see any signal in the centrosome. So now I don't know what I can do.
Does any one has any suggestion? Thank you!
Edited by Christina Shi, 24 November 2010 - 05:31 AM.