RE: IHC question
it always helps to put some normal serum from the animal you are staining
in the secondary mix. if you are staining human lung, put human serum in
your secondary, if you are staining sheep put sheep serum in your secondary,
about 20%, another thing and this does not make any sense, but DAKO told me
to use swine serum to block with even though their secondary was made in
goat, it worked, go figure.
From: Rebecca S Smith
To: HistoNet Server
Sent: 10/20/00 4:02 PM
Subject: IHC question
We have a project that is continually giving us background problems. At
low dilutions of normal rabbit serum used as a control (for hyper-immune
rabbit serum) in lung tissue from sheep and human, we have background
staining of respiratory epithelial cells. We have tried blocking with:
BSA, with 5% BSA with 20% normal goat serum(as that is the species of
secondary), with 5% BSA with 2% casein, and Biogenix powerblock.
Does anyone have any good ways to inhibit background staining of
rabbit serum in sheep and/or human lung?
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