[Histonet] Quick H202 quenching question.

[Lewis, Patrick]

If I have an IHC where I am staining 2 slides from the same block, one with one primary antibody and the other with a different primary antibody.

And one antibody's slides have high nonspecific background, but the other antibody's slides have no background,  Can I deduce that the background staining is not caused by insufficient H202 quenching/blocking?

All the steps were the same, except for the primary antibody/secondary antibody.  Also, they did have different epitope retrievals.

But the wash buffers/blocking buffers/and substrate were the same.

I am fairly confident that my background problems are related to this particular primary antibody, and not the actual quenching/blocknig from my IHC protocol, but I thought I'd check with you guys.

Patrick.

Patrick Lewis
Research Associate II Bench
Seattle Childrens Research Institute
206-884-1115

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