Hi Jim,
we use ice cold acetone for 5min. It seems to produce less background with our
antibodies and usually don't need an extra permeabilisation step. My experience
with 4% NBF was that the antibody thats supposed to stain intracellular vesicle
doesn't like it;-)
Good luck,
Frauke
--
Quoting Lee Crosby :
> Jim,
> I routinely stain cells with fluorescent antibodies. I fix them with 4%
> Paraformaldehyde in PBS, or the easier way, which I use now, is in neutral
> buffered formalin, which is readily purchased and does not require the labor
> intensive steps of the former.
>
> Good luck.
>
> Lee Crosby
> Echelon Biosciences Inc.
> 801-588-0455 ext 329
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vickroy, Jim
> Sent: Monday, November 03, 2008 12:58 PM
> To: 'histonet@lists.utsouthwestern.edu'
> Subject: [Histonet] CELL CULTURE LINE ON SLIDES
>
>
> We were asked to do immunostains on slides where a particular cell line was
> growing. Can anyone tell me the best way to fix these slides before
> performing immunostains? We used to use acetone on cytospins but can't
> recall if there is a better way.
>
> thanks
>
>
>
>
> Jim Vickroy BS, HT(ASCP)
> Technical Supervisor - Surgical and Autopsy Pathology
> Memorial Medical Center
> 217-788-4046
> vickroy.jim@mhsil.com
>
>
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