Re: Myeloperoxidase reference

From: Elizabeth Paietta (paietta@aecom.yu.edu)
Date: Fri Mar 21 1997 - 08:09:33 EST


On Wed, 19 Mar 1997, Ellen Ko wrote:

> 
> 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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