[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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Hello everybody,


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

following antibodies:


- 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..............


Richard  Edwards



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).


Good luck,


Kim Merriam'


Cambridge, MA



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.


Jan Shivers

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.

Best wishes,






I had problems years ago using an anti-rabbit secondary.  I'm attaching our




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