lysing solutions

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We have been using the CALlyse reagent for CD3 and CD19 enumeration of hu
WB using FITC conjugated MAbs with good results. However, when we use
CD3-biotin or CD19-biotin followed by Av-FITC, the histogram is a mess: the
fluorescence of the negative cells increases significantly and the CD3
positive cells are split in brights and dims.

So far we've tried extra washes after the Av step, but this did not help,
it seems that the Av is redistributed among the cells during the lysing
step.

Before trying the other commercial solutions, I'd like to hear from others
who have tried indirect labelling followed by a lyse/fix reagent.

Thanks in advance.

Jan.

Jan F. Keij
Los Alamos National Laboratory
Life Sciences Division
LS-5, Cytometry, MS M888
Los Alamos, NM 87544
USA

tel : (505)-667-3526
fax : (505)-665-6894
e-mail : keij@telomere.lanl.gov

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• Previous message: Dr. Howard Petrie: "SUMMARY: PI/DNA plus FITC/PE surface staining"

CD ROM Vol 2 was produced by 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 Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu EMAIL robinson@flowcyt.cyto.purdue.edu