Maurice R.G. O'Gorman, et. al. A rapid whole blood lysis technique for the
diagnosis of moderate or severe leukocyte adhesion deficiency (LAD).
Annals of the NYAS, vol. 677;427-430 (1993).
We ended up using PMA only (no ionomycin) and added it directly to whole
blood at an optimal final concentration of 120ng/ml and then measured the
Beta 2 leukocyte integrin expression by gating on grans, monos or lymphs
simply by their corresponding light scatter properties. Above this
concentration the density of the integrins on the cell surface continued to
increase but you begin to get considerable cell loss. We now routinely use
100ng/ml final (easy to make up) in our current clincal assay. If you
would like our laboratory procedure, we will fax it to you.
Jane Hughes wrote:
Dear Flowers
I am setting up an assay to screen for the expression of CD18,
11a,11b and 11c on neutrophils; taking mean peak intensity of
resting versus stimulated cells.
Is it possible that some of you have used Ca ionophore for this
purpose and could advise me as to an appropriate concentration fro
stimulation ( or advise a reference).
Many thanks
Jane Hughes
Immunology Section
Dept Paediatrics
Red Cross Children's Hospital
Rondebosch, Cape Town, South Africa