In common with many clinical flow'ers we routinely use Caltag Fix and Perm
reagent for MPO detection. It is true that most "in house" methods of
permeabilisation will work for MPO. We have tried various methods using
non-ionic detergents and commercial rbc lysing solutions. They all had a common
fault when using whole blood or bone marrow, positive MPO expression is
"induced" in otherwise negative cells by effusion from MPO positive myeloid
cells. Somehow the Fix & Perm reagents overcome this problem. If you are using
relatively pure cell suspension i.e. less than 5% contaminating normal myeloid
cells then most permeabilisation methods will work, if not I recommend Fix &
Perm.
Despite what Ellen Ko said, Fix & Perm works well for the detction of TdT
particularly if you use the HT6 clone, which is available from Supertech and
DAKO. Dr Elisabeth Paietta has published many papers on the subject.
Regards
*********************************************************************
Len Brown 102554.2147@compuserve.com
Department of Haematology (968)515736 voice
Sultan Qaboos University (968)513419 fax
Al Khod, Code 123, Sultanate of Oman