[Histonet] Re: rabbit anti-FITC

[C.M. van der Loos]

   Hi Liz, Gayle and Andrea,

   We were quite annoyed here too about discontinuing rabbit anti-FITC by
   Dako's. After complaining about it Dako let me know that a HRP-labeled
   rabbit  anti-FITC  is still  available.  However,  that approach isn't
   sensitive enough  for my application below. I looked for a replacement
   and found   a   concentrated   rabbit  anti-FITC  from  BioGenesis no.
   4510-7804 ([1]www.biogenesis.co.uk). This   product still   needs   to
   be compared with the original Dako rabbit anti-FITC. When done, I will
   let you know my findings. The test will be a
   multistep enzymatic double  staining  procedure combining an unlabeled
   mouse primary with a FITC-labeled mouse primary antibody:
    1. unlabeled mouse primary 1
    2. polymer anti-mouse/HRP
    3. blocking step with normal mouse serum
    4. FITC-labeled mouse primary 2
    5. rabbit anti-FITC
    6. goat anti-rabbit/AP
    7. visualization of AP activity (Fast Blue BB or Vector Blue)
    8. visualization of HRP activity in red (AEC or Vector NovaRed)

   In this protocol a mouse anti-FITC obviously doesn't fit.

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


   Date: Thu, 18 May 2006 16:10:05 -0600
   From: Gayle Callis <gcallis <@t> montana.edu>
   Subject: Re: [Histonet] rabbit anti-FITC
   To: "Elizabeth Chlipala" <liz <@t> premierlab.com>,
   Histonet <@t> lists.utsouthwestern.edu
   This  is sad as this antibody worked so nicely.  It worked beautifully
   with
   Rat antMouse CD4-FITC.  Chris van der Loos was the one who clued me in
   to
   this  antibody, as he used it cleverly for his double staining.  Maybe
   he
   has a new source for one of equal quality.
   Gayle Callis
   At 10:59 AM 5/18/2006, you wrote:
   >Hello everyone
   >
   >I was searching the histonet archives and came across an e-mail from
   >Gayle  Callis  about  the use of Dako's rabbit anti-FITC antibody for
   FITC
   >labeled  mouse  antibodies  on  mouse  tissue.   To make a long story
   short,
   >it  seems  that  Dako has discontiued this item, is anyone aware of a
   good
   >subsititue for this,  the one I was intere! sted in w

References

   1. http://www.biogenesis.co.uk/