[Histonet] RE: Biotin conjugated Antibodies -- can we skip biotin??

pruegg <@t> ihctech.net pruegg <@t> ihctech.net
Sun Jul 31 09:34:49 CDT 2005

Chris Is correct here I use what ever secondary I have that is against the
primary antibody species even if it is conjugated with biotin or fitc with a
labeled polymer hrp detection system to avoid the problem of unblockable
endogenous biotin in the tissue.  I just got a labeled polymer detection
system for primary antibodies made in goats from Invitrogen (I had trouble
finding one for goat primary antibodies).

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of C.M. van der
Sent: Wednesday, July 27, 2005 6:14 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Biotin conjugated Antibodies -- can we skip biotin??


   You may apply successfully any appropriate anti-goat reagent, ignoring
   the biotin-label of the primary antibody. Biotin groups at the primary
   are far too small to block anti-goat reagents. As long as there are no
   biotin-binding  structures  in  the section, which is only very rarely
   the  case, the biotin-label of the primary isn't influencing the final
   staining result.

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

   phone:  +31 20 5665631
   fax:    +31 20 6960389

   Date: Tue, 26 Jul 2005 12:23:10 -0700 (PDT)
   From: GT Hebert <emerald_lake77 <@t> yahoo.com>
   Subject: [Histonet] Biotin conjugated Antibodies -- can we skip
   To: histonet <@t> lists.utsouthwestern.edu
   Just a general question to help increase my knowledge:
   If I had an antibody that was conjugated to biotin -- say for example,
   a  goat  anti-mouse  P-selectin  (biotin)  -- can I just take a rabbit
   anti-goat  conjugated to AP and run IHC that way -- hopefully avoiding
   the  ABC  step  and  background OR does the biotin from the primary Ab
   block secondaries from coming in?
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