Re: [Histonet] different antigens need different methods for antigenretrieval?

From:"Katri Tuomala"

Generally speaking each antibody is optimized by the manufacturer (if not, 
it should be) as far as, application, antigen retrieval and approximate 
dilution. Their specification sheet gives you that information. That is your 
starting point to set it up in your detection system.
If your test material is outside the application recommended for that 
antibody, it still may work, but you will have to do a lot more trials, 
before you know that. For instance, if the application is said to be for 
frozen section, but you want to use it with formalin fixed paraffin embedded 
tissue, you'll have to do as complete work-up as you can. ie. try various=20
enzymatic digestions, HIER in different buffers or no retrieval at all. It 
can sometimes take days and even weeks of concentrated effort and even then 
you may not be able to make an antibody to work for something it was not 
meant for. So, when selecting an antibody, try to match it for the 
application you will need it for.
Hope this helps a bit...

 Katri Tuomala
Hamilton, Ontario, Canada

----- Original Message ----- 
From: "pex" 
Sent: Thursday, June 02, 2005 12:46 PM
Subject: [Histonet] different antigens need different methods for 

> Hello, all,
> I feel confused for immunofluorescence.
> I am doing immunofluorescence in vertebra sections. I use the same 
> protocol for immunostaining, but the results are very different, the one 
> is very nice, but the other is weak, I do not know the reasons. Do 
> different antigens need different methods for antigen retrieval?
> Generally, which methods for antigen retrieval in bone sections are 
> better? (Enzyme digestion? or HIER? or both?)
> Any suggestions will be helpful for me!
> Thank you!
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