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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