[Histonet] A multi-use simple blocking solution?....

[Patsy Ruegg]

For blocking in general I use a serum free protein block before the primary
antibody and then again before the secondary.  If I still experience
problems with non specific binding (usually of the secondary, but not
always), I will go more aggressive by using serum block matched to the host
of the secondary (usually a 10% solution without BSA) and if this still does
not do the trick I add to the serum block 10% normal sera from the species I
am trying to label. (Human serum is added to the serum block for human
tissue samples, mouse serum added to the block for mouse tissue samples,
etc.)  You must be careful not to block all of the antigen sites on your
tissue, though.
Patsy 


Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 216
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net
 

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gayle Callis
Sent: Friday, February 10, 2006 8:46 AM
To: Glenn Krasinski; Histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] A multi-use simple blocking solution?....

We prefer to use normal serum blocking, with the serum matched to the host
of the secondary and do not use BSA based blockers.  This has been discussed
at great lengths on Histonet, check out Histonet Archives.  Blocking, per
your question, is done many ways.

I suggest you do to the DAKO website, information and access the Handbook of
Immunochemical Methods, Boesnish discusses various blocking agents, and you
can download this wonderful freebie in pdf format.



You can buy superblocks ready to use from various companies, i.e.At 03:13 PM
2/9/2006, you wrote:
>I have been looking through the protocols on some of the IHC databases 
>for blocking solutions.  Some of the recipes are fairly exotic (fish 
>skin gelatin?).  Is there a good simple blocking solution that can be 
>used in a variety of applications (primary/secondary)?  How about 1x PBS
with 1% BSA?
>
>   Many thanks.
>
>   Glenn M. Krasinski
>   San Diego, CA
>
>
>---------------------------------
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)



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