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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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CD-ROM Vol 3 was produced by Monica M. Shively and other 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
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