AW: [Histonet] Chemicals that inactivate the primary antibody

[Merced M Leiker]

Hi TF and all interested,

I think I know what you want, but unfortunately I don't know how to answer 
your question (it is something I'd like answered myself!!)  To re-word for 
the sake of all interested:

You want to perform double-immunofluorescent staining using 2 primaries 
that were raised in the same species.

An additional question for clarification is: Do you want to do this on 
paraffin or frozen sections?

Maybe someone can help figure it out...

Merced


--On Sunday, May 17, 2009 2:16 AM +0800 TF <tifei <@t> foxmail.com> wrote:

> But the heat also damage the fluorescnece...
> u need specific amplication kit anyway.
>
>
> 2009-05-17
>
>
>
> TF
>
>
>
> 发件人： Gudrun Lang
> 发送时间： 2009-05-17  02:00:18
> 收件人： tifei <@t> foxmail.com
> 抄送： histonet <@t> lists.utsouthwestern.edu
> 主题： AW: [Histonet] Chemicals that inactivate the primary antibody
>
> For doublestaining the primary undergoes denaturation through a second
> HIER-step. The second secondary ab doesn't bind to the first primary.
> Therefore the binding sites must have been destroyed by heat in
> retrieval-buffer.
> Gudrun
> -----Urspr黱gliche Nachricht-----
> Von: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von TF
> Gesendet: Samstag, 16. Mai 2009 18:48
> An: Histonet <@t> lists.utsouthwestern.edu
> Betreff: [Histonet] Chemicals that inactivate the primary antibody
> Hi all, just wonder what kind of treatments/chemicals can complete block
> the binding of 2nd antibody to binded primary antibody on antigen?
> I tried HCl , but does not damage all the primaryantibody binding sites -
> i can still see staining pattern finally.
> 2009-05-17
> TF
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Merced M Leiker
Cardiovascular Medicine
348 Biomedical Research Building
State University of New York at Buffalo
3435 Main St, Buffalo, NY  14214
leiker <@t> buffalo.edu
716-829-6118

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