[Histonet] HRP-conjugated secondary and biotinylated-secondary

[Patsy Ruegg]

Subrata,
An advantage to using hrp conjugated secondary antibody instead of
biotinylated is that you can avoid the possible interference of endogenous
biotin which for some tissues may be difficult or impossible to get rid of
in my experience.  A disadvantage is that the indirect, two step hrp
conjugated method is not as sensitive as the ABC method (you build on to the
molecule by having more binding sites with the ABC method than with the hrp
conjugated method.  My choice for detection when possible is to use a
labeled polymer which is conjugated to hrp.  The sensitivity is as great or
greater as the ABC method without the endogenous biotin problem.
Patsy Ruegg

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Subratab
Sent: Saturday, January 29, 2005 11:19 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] HRP-conjugated secondary and biotinylated-secondary


   Hi,
   Is it a common practice to use HRP-conjugated secondary antibody
   instead of biotinylated secondary antibody for immunohistochemistry
   (with DAB developing system)? I am confused about the advantage/
   disadvantage of using HRP-conjugated secondary antibody for immunos.
   Subrata Biswas
   UNICAMP, SP, Brazil.
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