[Histonet] RE: True Blue chromogen

[C.M. van der Loos]

   Margaret,

   TrueBlue  yields  the  type  of color that is basically turquoise; one
   calls  it green and others blue. But indeed TrueBlue is more blue than
   green.  The  original protocol (also in the book under 'non-commercial
   visualization systems') using TMB chromogen is more greenish. However,
   both  TMB  and  TrueBlue  are  very  sensitive/efficient but difficult
   chromgens  in  terms of good staining intensity and low background. To
   my  experience  it  doesn't  always work for any antibody. Usually you
   have  to  dilute  your  primary  at  least 5-50x more than usual. Also
   dilution of your secondary (2-10x) is optional.

   Lots of success!

   Chris van der Loos, PhD
   Dept. of Pathology
   Academic Medical Center M2-230
   Meibergdreef 9
   NL-1105 AZ Amsterdam
   The Netherlands


   Date: Tue, 12 Sep 2006 11:27:54 -0500
   From: "Perry, Margaret" <Margaret.Perry <@t> sdstate.edu>
   Subject: [Histonet] True Blue chromogen
   To: <histonet <@t> lists.utsouthwestern.edu>
   I  am  in  need  of some clarification on information in Chris van der
   Loos
   book  "Immunoenzyme  Multiple  Staining  Methods".   On page 45 in the
   chart
   he  lists  TMB  HRP  substrate  as  producing a green color.  In KPL's
   catalog
   it lists TMB as blue and I talked to their tech service and they also
   said it turns blue.  Do you have to do something to the TMB to turn it
   green or does it turn green automatically with IHC ?  Are there any
   other chromogens that turn green?

   Thanks in advance for your help.

   Margaret Perry HT(ASCP)
   South Dakota State University
   Vet Science