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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