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>
> 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
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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
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Phone: (765)-494-0757;
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