Rabbit antibody on rabbit tissue

From:"Mark Elliott"

Hi everyone. I am currently trying to use a rabbit polyclonal antibody
to endothelin-1 (Peninsula labs) on formalin-fixed, paraffin-embedded
rabbit tissue and of course am getting a lot of background.  I have used
the antibody on human and pig tissue with no problems.  Looking at the
methods for doing mouse on mouse I tried adding the primary and
secondary (biotinylated goat and rabbit)  together at the same
concentrations as used separately before adding it to the slides.  I
have also tried using higher concentrations of primary, or lower
concentrations of secondary in combing the two antibodies but in all
cases there is no labelling what so ever on either rabbit tissue or my
normal human positive control tissue.  I have added rabbit IgG to the
primary and secondary mix as well with no effect.  Does anybody have a
method of using rabbit polyclonals on rabbit tissue they care to share. 
Or does anyone know of a mouse anti-endothelin-1 that would work on
formalin fixed paraffin-embedded rabbit tissues?
Thanks for any input anyone can give.


W. Mark Elliott, PhD
Research Associate

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