[Histonet] summary of reactions to porcine insulin
Stryker, S. (HLK)
S.Stryker <@t> lumc.nl
Thu Apr 8 02:43:15 CDT 2004
summary of reactions to:
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I'm hoping someone can give me any suggestions on how to detect insulin on a
(paraffin embedded formalin-fixed or frozen) porcine section using the
- primary antibody: Guinea Pig anti-insulin (Dako)
- secondary antibody: Swine anti-Rabbit (Dako)
According to the DAKO catalogous the anti-Rabbit should work in this combo.
I have tried it over and over again, using different protocols, but somehow
I just can't get the job done...
Thanks in advance!
It will never work with this combination of antibodies, what you
need(and what I use)is a rabbit ANTI-GUINEA PIG peroxidase conjugated,
from DAKO(P0141) as your second antibody..............
would use a biotinylated anti-guinea pig antibody. I'm not sure if the
anti-rabbit antibody would bind the the GP primary antibody you are using.
I'm pretty sure that Vector sells an anti-GP secondary antibody (I think
they also sell a guinea pig ABC and alk phos kit, depending on the detection
system you are using).
es, a biotinylated anti-rabbit IgG does cross-react with guinea pig
antibodies. I use it all the time when I stain for Insulin, using Dako's
guinea pig anti-Insulin product.
U of MN Vet Diag Lab
I have had great success with that antibody.
I use it @1:100. I use Goat x Rabbit/Biotinylated @1:100.
I run this on the Ventana but you should get similar results maually.
Poly-HRP goat anti-rabbit IgG would react well with guinea pig primary
antibody. Kilinipath at Netherlands should be able to help. Klinipath:
31-316-250309 attention Marc or Dick.
I had problems years ago using an anti-rabbit secondary. I'm attaching our
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