rabbit on rabbit Re: [Histonet] endothelial cell specific lectin staining

From:Gayle Callis

Mark and Jan,

There is an excellent, extensive review of Lectins in J Histochemistry 
Cytochemistry if you need a place to start on lectin learning.  Check out 
this publication in the same journal, Glycoconjugate expression defines the 
origin and differentiatian pathway of intestinal M cells.  Geber A, Posselt 
W. 45(10):1341-1350, 1997.  This work was done on rabbit tissue.

Doing rabbit on rabbit IHC is very much like doing doing mouse on mouse 
IHC.  If one biotinylates the primary and then does the proper normal serum 
block success may be possible, similar to the mouse on mouse kits using 

There is a publication from a defunct journal,  Cell Vision by Gu J et al, 
titled a Primary secondary antibody complex method of immunocytochemistry 
using rabbit polyclonal antibodies to detect antigens in rabbit tissue, 
2(1):59, 1995.  This journal became Applied Immunohistochemistry and 
Molecular Morphology later on, and they may be a source of the article 
since Gu is one of the editors for AIMM.   You can also contact me 
personally if interlibrary loan fails or you can't locate the article.

The method is also in a book titled Analytivcal Morphology, Theory, 
Applications and Protocols, Ed. Jiang Gu, Chapter:  Elimination of 
Background Staining, using the primary secondary antibody complex 
method.  ISBN#1-881299-03-1, soft cover Eaton Publishing 1997 unless out of 
print.   Topics included FISH, immunogold, antigen retrieval, MW, etc).

but At 04:38 PM 11/27/2006, you wrote:
>Just my two cents worth... from my experience with lectins a couple 
>decades ago, I found that what worked in one species did not always work 
>in another... sometimes there was similarity between species, but not 
>always. So, if you choose to go the lectin route, do a good literature 
>search first or be prepared to run many trials.
>The cellular glycoconjugates (to which the lectins bind) may not be 
>exactly the same from species to species.  That was my guess at the time, 
>But, I agree... avoid rabbit antibodies on rabbit tissues, if at all 
>possible.  I've never been able to get decent staining without background 
>when using unconjugated polyclonal antibodies on rabbit tissues.
>Jan Shivers
>Univ. of Minnesota Vet Diag Lab

Gayle Callis HTL, HT, MT(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717

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