[Histonet] RE: IHC antibody optimizing & validating

[Cartun, Richard]

I have not read the entire document yet.  What do they say about using tissues that have been fixed and processed elsewhere?

Richard

Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 972-1596 Office
(860) 545-2204 Fax
richard.cartun <@t> hhchealth.org

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Terri Braud
Sent: Tuesday, March 25, 2014 3:35 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: IHC antibody optimizing & validating

CAP is very clear that in order to validate a new antibody, that: "once the stain has been optimized, that for a well characterized antibody with a limited spectrum of antigenic targets, like Chromogranin or PSA, the validation can be limited.  A panel of 10 positive and 10 negative neoplasms would be sufficient in this setting.  For an antibody that is not well characterized and/or has a wide range of reported reactivity, a more extensive validation is necessary.  The number of tissues tested should in the circumstance be large enough to determine whether the staining profile matches that previously described.  An exception to the above requirements is that studies nay not be feasible for antigens such as ALK that are only seen in rare tumors."
Thus sayeth CAP.
And if you're like me, I am not digging through all my cases to try to come up with 30-40 neoplasms for each antibody, so I just order Tissue Micro Arrays with the neoplasms I need.  20 positive and 20 negative neoplastic tissues on one slide for easy staining and validation.  The money you save on reagents to stain one little slide, more than makes up for the cost of the slide.
I hope this helps.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874

Today's Topics:
   8. IHC antibody optimizing & validating (Davis, Cassie)

Message: 8
Date: Tue, 25 Mar 2014 09:51:07 -0400
From: "Davis, Cassie" <CDavis <@t> che-east.org>
Subject: [Histonet] IHC antibody optimizing & validating

Will you help me? I understand we are to use the known positives controls that the manufactures' recommends in the package insert when optimizing the stains, but I need to know what is your general procedure for optimizing (how many different staining protocols do you test) and validating a new antibody (how many different or "known" positive and negative tissues do you test [predictive markers I understand are 20])?

Cassandra Davis
CDavis <@t> che-east.org
302-575-8095

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