Immunohistochemistry Puzzler

From:
Elizaha@aol.com

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Hi,
     There is an odd problem with an antibody we are using in our lab. It is 
a rabbit polyclonal, we use a streptavidin/biotin (commercial) kit, and 
chromagen is DAB. Tissue: chondrocytes/cell culture fixed in paraformaldehyde 
and embedded in paraffin..
Problem:  When we stain cells with it we get little to no label, if we add a 
hydrogen peroxide blocking step before the primary antibody we get heavy 
labeling of cells that looks specific/intracellular. The antibody is made "in 
house" but is not against hydrogen peroxide? Has anyone seen any reaction 
like this. Is this weird or what?

Thanks again, 
Elizabeth

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<HTML><FONT FACE=arial,helvetica><FONT  SIZE=2>Hi,
<BR>     There is an odd problem with an antibody we are using in our lab. It is 
<BR>a rabbit polyclonal, we use a streptavidin/biotin (commercial) kit, and 
<BR>chromagen is DAB. Tissue: chondrocytes/cell culture fixed in paraformaldehyde 
<BR>and embedded in paraffin..
<BR>Problem:  When we stain cells with it we get little to no label, if we add a 
<BR>hydrogen peroxide blocking step before the primary antibody we get heavy 
<BR>labeling of cells that looks specific/intracellular. The antibody is made "in 
<BR>house" but is not against hydrogen peroxide? Has anyone seen any reaction 
<BR>like this. Is this weird or what?
<BR>
<BR>Thanks again, 
<BR>Elizabeth</FONT></HTML>

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