RE: elastase in neurtophils

From: Gregor Rothe (gregor.rothe@klinik.uni-regensburg.de)
Date: Mon Apr 10 2000 - 02:59:05 EST


You should be at least extremely careful when attempting to analyse a
specific enzyme with Cell Probe reagents. In contrast to our previously
published work (Biol Chem Hoppe-Seyler 373: 547-54, 1992) on the
intracellular analysis of elastase with rhodamine 110 based substrates, many
Cell Probe reagents are not blocked against non-specific exopeptidase
activity. Also except citing the specificity of an analog non-rhodamine
based substrate in a cell lysate assay, no data are given about the
specificity of the reagents in an intracellular assay. This information
could be easily achieved using cell-permeable and specific inhibitors. I am,
however, not aware of any such data for Cell Probe reagents.

Gregor Rothe

> -----Original Message-----
> From: Christina McCowan [mailto:c.mccowan@pgrad.unimelb.edu.au]
> Sent: Friday, April 07, 2000 1:43 AM
> To: Cytometry Mailing List
> Subject: Re: elastase in neurtophils
>
>
>
> Virginia,
>
> You might like also to check out the Cell Probe reagents marketed by
> Beckman Coulter.
>
> http://134.217.3.35/coulter/cytometry/CellProbe-Reagents/cp-index.asp
>
>
> Christina McCowan
>
>
> At 18:49 05-04-2000 -0500, you wrote:
> >
> >Virginia,
> >
> >I have not tried this; but, you micht consider looking at this
> possibility.
> >http://www.phiphilux.com/elas1.htm
> >I think this is suppose to be a cell permeable substrate that changes
> >fluorescent properties upon protease cleavage.
> >Ty Lee
> >
> >
> >-----Original Message-----
> >From: Virginia M Litwin <virginia.litwin@bms.com>
> >To: cyto-inbox
> >Date: Wednesday, April 05, 2000 5:12 PM
> >Subject: elastase in neurtophils
> >
> >
> >>Does anyone know of a method to measure elastase in neutorphils?
> >>
> >
>



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