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
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:33 EST