[Histonet] antibody validation Thanks, but need Non-"U" also

[Akemi Allison-Tacha]

Hi All,

I would like to thank those of you who have responded to the e-mail below regarding antibody validation.  The information I received was pretty much what I expected from university IHC labs.  I would like to know what smaller IHC labs are doing for validation and re-validation when changes are made to their protocols.

Thank you again for any and all responses,
Akemi

Akemi Allison-Tacha BS, HT(ASCP)HTL
Histology Manager
APMG Laboratories
105A Cooper Court, Los Gatos, CA 95032
Contact: 800.848.2764
V/M: 408.884.2718
Fax: 408.884.2758
Cell: 408.335.9994
(W) E-Mail: aallison-tacha <@t> apmglab.com
(P) E-Mail: akemiat3377 <@t> yahoo.com



--- On Fri, 9/11/09, Akemi Allison-Tacha <akemiat3377 <@t> yahoo.com> wrote:

From: Akemi Allison-Tacha <akemiat3377 <@t> yahoo.com>
Subject: [Histonet] antibody validation
To: "histonet" <histonet <@t> lists.utsouthwestern.edu>
Date: Friday, September 11, 2009, 8:20 AM

Good Morning and Happy Friday out there in Histo-Land!

I would like your assistance in an issue that I have just become aware of regarding antibody validation.  I have been requested to bring on-board approximately (7) new antibodies in the immediate future, and several antibodies later.

Although, many of you who know me realize I have worked in IHC R&D for a well known IHC company for many years, it has been a while since I have overseen a clinical IHC lab.  I would like your assistance and input on how you are validating new antibodies.   Not counting Her2-neu, it has been my understanding that it is recommended to test samples with varying antigen expression from 10-15 cases.  Furthermore, you should check for Sensitivity: expression range of Positive cases, low to high (10-15 cases).  Specificity: (+) vs expected (-) cases/tissues (10-15 cases) and Precision (Reproducibility): 

It has been my practice that any changes that are made to existing protocols, such as a new detection kit or antigen retrieval methodology, also requires re-validation.  

I am curious how you approach validation.  Also, do any of you just use a single known positive control for that antibody, and run it, and if it is positive, feel that it is satisfactory, and put it on-line for testing.

Thank you in advance for your input, and have a great weekend!

Akemi Allison-Tacha BS, HT(ASCP)HTL
Histology Manager
APMG Laboratories
105A Cooper Court, Los Gatos, CA 95032
Contact: 800.848.2764
V/M: 408.884.2718
Fax: 408.884.2758
Cell: 408.335.9994
(W) E-Mail: aallison-tacha <@t> apmglab.com
(P) E-Mail: akemiat3377 <@t> yahoo.com

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