From: William Ainsley Baughman (wab@gwis2.circ.gwu.edu)
Date: Fri Oct 06 1995 - 17:24:12 EST
Caltag fix/perm "kits" work fine. Some loss of SSC is generated by the perm reagent. We have used the MPO/Lactoferrin, MPO/CD3, and MPO/CD22 cocktails by Caltag. These cocktails also work with BD's FACSLyse reagent. BD's anti-CD3 conjugates stain cytoplasmic antigens well. Kirkegaard-Perry's anti-kappa or -lambda stain cytoplasmic antigens well, while the surface has been tagged with CD38(PE) and CD45(CY5-PE). We have not observed cytoplasmic CD22 after the surface has been found to be negative. I don't know of other mfgs of anti-MPO. NH4Cl treated specimens tend to have more loss of SSC than whole blood. Even if the SSC is pushed up to match lymphocytes (or blasts) for region analysis, separation (with regard to light scatter) between myeloids and monos and blasts and plasma cells and whatever else is there is less. ====================== | Bill Baughman | | 703-802-7070 x5807 | ====================== On 6 Oct 1995, LEONARD BROWN wrote: > Hi > > Does anyone have experience with intracellular staining of whole blood using MPO > MoAbs. We are unable to overcome induction in MPO negative cells with samples > having high neutrophil counts. Have tried lots of permeabilising agents and > fixation strengths. Has anybody used 'Fix and Perm' from Caltag and does it > really work for MPO using whole blood methods. > > > Len Brown 102554.2147@compuserve.com > > Department of Haematology (968)515736 voice > Sultan Qaboos University (968)513419 fax > Al Khod, Code 123, Sultanate of Oman >
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