BrdU Staining

Paul Zimmer Paul.Zimmer at
Thu Feb 27 17:11:48 EST 1997

Our laboratory has been using the standard acid-denaturation protocol
for testing BrdU incorporation by flow, but we have been disappointed by
the number of common epitopes destroyed during this process (e.g. CD23
on murine B-cells).  We have been testing the DNAse protocol described
by John Sprent's group, but with little success.  All of the key
reagents were recently purchased and work well in other assays.  I am
curious about two aspects of the protocol:

1) Does azide interfere with the DNAse enzyme activity? (we use azide in
our common stock of 1% paraformaldehyde solution)

2) Does the source of DNAse make a difference? (we have been using
molecular biology grade RNAse-free DNAse I)

Any other suggestions or helpful comments on common problems would be
greatly appreciated.  We have already spoken with Dr. Sprent's lab and
they have not had similar problems.

J. Paul Zimmer, Ph.D.
Developmental and Clinical Immunology
University of Alabama at Birmingham

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