Preabsorbed polyclonals

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From:David_Howlett-1@sbphrd.com
To:histonet@pathology.swmed.edu
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Does anyone know why preabsorbed antibodies sometimes give enhanced staining by
IHC?
I am working with an affinity purified antiserum, raised to a small peptide
fragment, which appears to give a reasonable staining pattern using Vector ABC
and DAB systems on paraffin sections of a variety of human and rodent tissues.
The preimmune serum produces very little staining.  However, preabsorbing the
antiserum with the peptide that it was raised against (antiserum and peptide
incubated together, various times, various temperatures) always gives more
intense overall staining on comparable sections.  I have occasionally noted this
with other antisera.
Any ideas?
Thanks
David Howlett
SmithKline Beecham Pharmaceuticals.





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