Re: Immunos and Oven Temps/Times

<< Previous Message | Next Message >> (Elizabeth Morehead),
Date:Mon, 19 Apr 1999 11:15:19 -0400 (Eastern Daylight Time)


Don't know about this "Guru" stuff but will share what little I know what has worked for me ;-).

Once the slides are cut  and put on + slides (Fisher) I have the histology lab let the slides air
dry overnight, NO slidedryer.  While this is not necessary for the demonstration of many epitopes it
IS  for some of the ones we routinely look at.  

For MOST of our primary antibodies the incubation time is set at 10 (sometimes 15) minutes at room
temp., using the Dako Envision Kit (HRP) for the demonstration system, which means that the
secondary conjugated antibody is also a 10 min. RT incubation.  

There are of course exceptions (aren't there always) which makes life interesting.  We have some
primaries which stain optimally only with 37oC incubation for 1 to as many as 4 hours and some that
required overnight at roc incubations.  But those are the longest incubation times I have ever used.

I must admit that 48+ hours at 70oC seems a tad bit excessive.

-- Begin original message --
> How long are you incubating your immuno slides?  What temp?
> Anybody ever heard of leaving immuno control slides in a 70 degree oven for the weekend?  Yes, I 
> said WEEKEND!   Anybody doing anything special to keep fat on slides (dermis and breast)?
>  Your answers could very possibly win (or lose) me a bet and your opinions are most appreciated!  
> Thanks in advance.
> Beth
> Beth Morehead
> Med U of SC
> (843) 792-4223
-- End original message --

best regards,
Robert Schoonhoven
Laboratory of Molecular Carcinogenesis and Mutagenesis
Dept. of Environmental Sciences and Engineering
University of North Carolina
Chapel Hill, NC 27599
office 919-966-6343
   Lab 919-966-6140
   Fax 919-966-6123 

**Suppose you were an idiot... And suppose you were a member of Congress ...
But I repeat myself.-Mark Twain**

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