[Histonet] different antigens need different methods for
katri <@t> cogeco.ca
Thu Jun 2 19:10:04 CDT 2005
Generally speaking each antibody is optimized by the manufacturer (if not=
it should be) as far as, application, antigen retrieval and approximate=20
dilution. Their specification sheet gives you that information. That is y=
starting point to set it up in your detection system.
If your test material is outside the application recommended for that=20
antibody, it still may work, but you will have to do a lot more trials,=20
before you know that. For instance, if the application is said to be for=20
frozen section, but you want to use it with formalin fixed paraffin embed=
tissue, you'll have to do as complete work-up as you can. ie. try various=
enzymatic digestions, HIER in different buffers or no retrieval at all. I=
can sometimes take days and even weeks of concentrated effort and even th=
you may not be able to make an antibody to work for something it was not=20
meant for. So, when selecting an antibody, try to match it for the=20
application you will need it for.
Hope this helps a bit...
Hamilton, Ontario, Canada
----- Original Message -----=20
From: "pex" <pex0220 <@t> yahoo.com.cn>
To: <Histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, June 02, 2005 12:46 PM
Subject: [Histonet] different antigens need different methods for=20
> Hello, all,
> I feel confused for immunofluorescence.
> I am doing immunofluorescence in vertebra sections. I use the same=20
> protocol for immunostaining, but the results are very different, the on=
> is very nice, but the other is weak, I do not know the reasons. Do=20
> different antigens need different methods for antigen retrieval?
> Generally, which methods for antigen retrieval in bone sections are=20
> better? (Enzyme digestion? or HIER? or both?)
> Any suggestions will be helpful for me!
> Thank you!
> DO YOU YAHOO!?
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