Re: Cell Culture for IHC

From:Edda Stoinski

You can use a formalin fixation for cell clutures, but not longer as 2 
minutes. Then you don#180#t need a retrival. Better is PFA buffered in PBS 
pH7.4. I do that with selfmade antibodys and i have best results. For 
cultivation we are used poly-L-Lysine coated cover glass slides, after 
the coating rinse with PBS . Chamber slides are also very good but to 
much expensive, in normal cover glass is ok. Don#180#t use a detergenz 
(Tween or triton) it disolved your cells from the glass surface. You can 
also use a petri dish for cultivate the cells,but then you need more 
antibody solution but it works good when you drawn a line arround a good 
area with a wax pencile.
Edda schrieb:

>I have been asked to do an immunohistochemistry stain (West Nile Virus) on cell culture.  I usually on worked with paraffin blocks and smears.  I'm not even sure of the questions to ask, but here it goes....
>In growing the cells, can I used glass slides (I don't have any plastic)?
>Are the glass slides plain or charged?
>How do I fix the cells?  (I know if they're not fixed in formalin there will be no antigen retrieval.)
>Anything special I need to know before the jump such as potential problems or background staining?
>Thanks in advance,
>Laura Bliven
>Marshfield Laboratories

<< Previous Message | Next Message >>