[Histonet] avian/biotin block

[Patsy Ruegg]

Has anyone tried the Signet Acuity Mouse-on-Mouse Detection System "Biotin
Free"?  This might be a good alternative to an ABC detection for MonM here.
Signet sent me a sample but I have not yet had time to try it.  It uses an
hrplabelled polymer link which is anti-ms and anti-rabbit with special
blocking reagents before the primary and another block before the link. 
Patsy

-----Original Message-----
From: anh2006 <@t> med.cornell.edu [mailto:anh2006 <@t> med.cornell.edu] 
Sent: Thursday, February 03, 2005 1:40 PM
To: Patsy Ruegg
Cc: 'Emerson, Rachael'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] avian/biotin block

Good idea - the polymers are superb - but EnVision is not suitable for mouse
embryos but you could look for one that was suitable.
I use DAKO Biotin Blocking System to routinely stain cell surface antigens
in mouse tissue even finicky hematopoietic tissues.
IMO, it is likely though that if your signal disappeared after blocking that
it was non-specific - do you have a positive control tissue where it
remained after blocking?



> I would consider using a non-biotin detection system such as Hrp 
> Labelled Polymer, there are several out there now, the one I use is 
> from DAKO called Envision.
> Patsy
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu
> [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of 
> Emerson, Rachael
> Sent: Thursday, February 03, 2005 12:32 PM
> To: 'histonet <@t> lists.utsouthwestern.edu'
> Subject: [Histonet] avian/biotin block
>
> Let me try to sum up a long story and get to my question......
> Basically I am staining mouse embryos-frozen sections, for megakaryocytes.
> I had trouble with endogenous AP and 15% acetic acid seemed to help. I 
> am having trouble with endogenous biotin, as well.  I tried Vector's 
> Avidin/Biotin Blocking Kit and lost my signal all together.  I am 
> looking for a surface antigen and I am afraid the kit blocked it all.
>
> I would really appreciate any suggestions on blocking endogenous 
> biotin on frozen sections?
>
> Also, I tried to us an AP secondary, but my signal was dimished.
> Thanks
> Rachael
>
> Rachael L. Emerson
> Center for Human Genetics and Molecular Pediatric Diseases University 
> of Rochester Medical Center
> 575 Elmwood Avenue MRBX 1.11301
> Rochester, NY 14642
>
> Tel (585) 275-5073
> Fax (585) 276-0232
>
>