[Histonet] RE:Immunofluorescent foci moving?

[Martin S.]

I used to have this problem when staining transcription sites in the
nuclues of cultured cells - we thought it was just clumps of antibody
getting trapped. I have found that FITC labeled antibody preps often
need to be centrifuged as some of the FITC seems to come out into the
solution. We centrifuged our antibody dilutions before use and increased
the number and length of washes and added 0.1% triton to the washes.
This seemed to help but occaisionally I still saw some moving foci so if
you get any interestng replies please let me know.
 
Best of luck
Sonya