Re: [Histonet] citation for negative controls

From:Gayle Callis

Appropriate negative controls are discussed by Boenisch, in DAKO 3rd 
Edition Handbook Immunochemical Staining methods, and found on the DAKO 
website as a free pdf download.  It can be done and clinical laboratories 
do use irrelevant antibodies at times also.  Jules Elias books also discuss 
this, Immunohistopathology, a pracatical approach to diagnostics, ASCP 
press. There is a second edition although may not be available to you.

If you are working with mouse, this can be a difficult thing to find at 
times so that you get NO staining with an irrelevant 
antibody.  Consequently we use what is most cited in the literature, 
isotype matched immunoglobulin i.e. IgG controls depending on species and 
immunoglobulin.  They come pure, biotinylated, conjugated to many things 
including Alexa dyes.  OR you can buy whole IgG's from Jackson (whole rat 
IgG contains all the isotypes, and from many different species.)

One that happened some time ago, we used rat serum as a  control and the 
reviewers balked big time.  We had to REPEAT what was a very difficult, 
tedious immunogold staining project using isotype matched controls.   Since 
that time, we NEVER use normal serums nor an irrelevant antibody for a 
negative control.   Having to repeat work is not fun!

  At 04:34 PM 2/1/2006, you wrote:
>I use IHC in basic research and have been digging through the archives for 
>information on appropriate negative controls and came across the use of 
>irrelavent antibodies. I would like to use this information in a paper I 
>am writing and was wondering if this is cited in a review or paper. I have 
>Harlow and Lane's book but it doesn't mention this type of control.
>Thank you in advance for any help.
>Jonathan Wilson (PhD)
>ecofisiologia CIMAR
>Rua dos Bragas 289
>4050-123 Porto Portugal
>office 351 22 340 1809
>lab 351 22 340 1834
>fax 351 22 339 0608
>Histonet mailing list

Gayle Callis HTL, HT, MT(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717

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