>
> A little info from some of our people in Germany:
>
> Regarding MPO:
>
> A. Fix & Perm is not the only way, but it is a good and easy way (it is
> often recommended in consensus protocols)! You can buy it from caltag
> in U.S. Most of our customers use this reagent! But it wont work for TdT
> staining!!
>
> B. MPO staining should also work by "handmade" methods:
> a) e.g., Paraformaldehyde / Ethanol (PFE) treatment; 2x10to 6 cells were
> fixed in 1 ml 1%PFA for 15 min.at RT. After that 1 ml PBS/BSA1% were added
> drop by drop. Cells then were pelleted and resuspended in 2 ml 45% ethanol
> in
> PBS and incubated for 30 min. at RT. Then cells were washed twice with
> PBS/BSA1% and stained.
> - this method should also work, but I do not have any experiences about
> background staining.
>
> C. An easy way for reducing background staining is to include a
> pre-incubation step with a mouse isotype-control antibody, not
> fluorochrome conjugated for 15 min. before the "real" staining starts. Some
> of our
> customers do this in case of Ki-67 staining.
>
> Jessica
> DAKO GmbH
>
>
> Ellen Ko
> DAKO Corporation
> Technical Service Specialist
> http://www.dakousa.com
> (800)424-0021
>
> ----------
> > From: Elaine Kunze <mek4@psu.edu>
> > To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
> > Subject: Myeloperoxidase reference
> > Date: Tuesday, March 18, 1997 6:48 AM
> >
> >
> > Myeloperoxidase reference:
> > "Flow cytometric detection of intracellular antigens for
> immunophenotyping
> > of normal and malignant leukocytes" K Groeneveid, et al. Leukemia
> (1996)
> > 10, 1383-1389.
> >
> >
> > ************************************
> > Elaine Kunze
> > Flow Cytometry......Image Analysis
> > The Biotechnology Institute for Research and Education
> > 8B Althouse Lab
> > Penn State University
> > University Park, PA 16802
> > (814) 863-2762
>
Just a correction of above message: Caltag's Fix & Perm works very well
for TdT in ALL as well as AML - we have published this: Leukemia and
Lymphoma, Vol 22:279-285, 1996.
Regards, E. Paietta