Comment on Ellen Ko's message

Walter Sharp (denby@compuserve.com)
Fri, 21 Mar 1997 03:27:50 -0500

I was going to reply "privately" but feel that others may want to comment
on my comments !

It seems strange to me that the talk about MPO "background staining" is
being looked at as simply that.
Most other commercial and in house methods are actually INDUCING the
activity by releasing the enzyme from Neutrophil lineage cells which then
gets drawn into the target population.
No amount of blocking will help in this case.
How An der Grub "prevent" this I do not know but it may have something to
do with the tremendous amount of "autofluorescence" seen after
permeablisation compared to, say, a Triton method.
Maybe a sort of covering-up effect.
Nevertheless, it does seem to work consistently with MPO.
As for TdT - I would argue the point that it doesn't work at all but I
would be the first to agree that you can get better results with harsher
permeablisation methods.
Just in case anyone thinks I'm "having a go" at Dako - we use their
excellent anti MPO and the HT6 TdT clone marketed by them (? Supertech in
the states).

Wal Sharp
SQU
Oman.


Home Page Table of Contents Sponsors E-Mail Archive Web Sites

CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone: (765)-494-0757; FAX(765) 494-0517; Web http://www.cyto.purdue.edu , EMAIL cdrom3@flowcyt.cyto.purdue.edu