[Histonet] Secondary Antibodies dilutions

Dardo Ferrara dardofer <@t> hotmail.com
Thu Aug 5 09:12:29 CDT 2004

I have had problem with biotinylated Abs .I have used 10ng/ml and it seems 
to be too much based on a lot of background in addition to some nonspecific 
binding even when using streptavidin conjugates...

Dardo Ferrara
Emory University

>From: Gayle Callis <gcallis <@t> montana.edu>
>To: Laboratorio de Histología 
><histolog <@t> fcv.unl.edu.ar>,Histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] Secondary Antibodies  dilutions
>Date: Tue, 03 Aug 2004 07:59:31 -0600
>A rule of thumb is to use 2 - 10 ug/ml, do the calculation from 
>concentration of antibody found as mg/ml in the specification sheets.  Some 
>people even use a lesser concentration - we have done dilution panels on 
>our secondaries at times to determine the optimal concentration needed in 
>ug/ml.   We tend to average around 1:250 dilution on 0.5mg/ml 
>concentration.  Specification sheets usually give the recommended dilution 
>for immunohistochemistry usage.
>At 05:41 PM 8/2/2004, you wrote:
>>I wanted to know if some of you uses the following  Antibodies  , and to
>>that dilution uses them in immunohistochemistry.
>>Biotin-Goat anti-Rabbit IgG (Zymed 65-6140
>>Biotin-Goat anti-Mouse (Chemicon AP181B)
>>Dr. Hugo H. Ortega (DMV, PhD)
>>Departament of Cellular Biology
>>Faculty of Veterinary Sciences
>>Universidad Nacional del Litoral
>>R.P. Kreder 2805 - Esperanza (3080)
>>Santa Fe - ARGENTINA
>>Tel. (54)3496-420639
>>Fax. (54)3496-426304
>>  <http://fcv.unl.edu.ar/histolog/> http://fcv.unl.edu.ar/histolog/
>>  <http://fcv.unl.edu.ar/bioterio/> http://fcv.unl.edu.ar/bioterio/
>>Histonet mailing list
>>Histonet <@t> lists.utsouthwestern.edu
>Gayle Callis
>Research Histopathology Supervisor
>Veterinary Molecular Biology
>Montana State University - Bozeman
>PO Box 173610
>Bozeman MT 59717-3610
>406 994-6367 (lab with voice mail)
>406 994-4303 (FAX)
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu

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