Mouse bone marrow protocol

From: Kimmo Porkka (kporkka@burnham-inst.org)
Date: Thu Mar 16 2000 - 10:38:33 EST


Hi!

I've been trying to devise/find a simple protocol for mouse bone marrow, but
so far with minimal success. I have not been able to get consistently clean
negative controls (either isotype matched or with second antibody only). I'm
working predominantly with biotinylated antibodies and FITC, PE or Cy-Chrome
labelled secondary reagents. I've tried different serum blocks (changed
concentration and species), Fc receptor block, lysed RBCs or not lysed,
titrated primary antibodies and excluded dead cells with PI or 7-AAD. My
purpose is to analyze rather rare populations in bone marrow (<1%) and that
is why my negative controls really should be negative.

Any help would be greatly appreciated.

Best regards,




Kimmo Porkka
Burnham Institute, La Jolla



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