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> Who does know any suitable protokol to measure elastase activity in
> leukocytes in FACS?
> Ulrich Sack
>
Human neutrophilic granulocyte elastase activity can be measured
kinetically intracellularly in viable cells using
(Z-Ala-Ala)2-Rhodamine 110. We published a protocol on the
the substrate which to my knowledge by now
is available from Molecular Probes in
Biol. Chem. Hoppe-Seyler 373: 547-554, 1992.
An analytical problem for flow cytometry, however, is that neutrophils
in suspension in contrast to adherent cells do only degranulate
to a maximum of minus 30% wether using the substrate or
anti-elastase antibodies. Therefore, in my opinion the substrate
is only of limited value at least in the setting of the analysis
of blood neutrophils in human sepsis.
Gregor Rothe
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Institute for Clinical Chemistry and Laboratory Medicine
University of Regensburg
D-93042 Regensburg, F.R. Germany
Tel. +49 (941) 944-6204
Fax +49 (941) 944-6202
Internet: Gregor.Rothe@klinik.uni-regensburg.de
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