Re: [Histonet] IHC on cell cultures

From:Sharon Cooperman

If the cells grow in a monolayer, just throw in a few cover slips 
(you may or may not have to treat the cover slips to get the cells to 
stick, depending on the cells) and then fix with whatever you like 
(4% PF in NBS, ice cold methanol, acetone, etc.).  If the cells grow 
in spinner culture, I think the best method is to do a cytospin.

Sharon

>Can anyone help me with a protocol for preparing cell culture samples for
>IHC?  I'm particularly interested in looking at CD45.  I'm not quite sure
>of the best way to prepare the cells.  I can do cytospins - and I'll bet I
>have to fix the cells prior to cytospinning them.  If anyone has a
>protocol they'd like to share for preparing good slides from live cells,
>I'd appreciate it.  I'd like to keep the option of using cytospins or
>smears as well as making a cell block.  I'm experienced with making these
>preparations from body fluids, but not cell cultures.  Thanks.
>
>Jackie O'.
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>Histonet@lists.utsouthwestern.edu
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-- 
Sharon Cooperman       	     
NIH, NICHD, CBMB                     301.435-8417
Building 18T, room 101               301.402-0078 fax
Bethesda, MD 20892

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