Re: B-lactamase assay in individual bacteria?

From: Phil Marder (mr_redram@hotmail.com)
Date: Wed Jun 07 2000 - 08:04:50 EST


The supplier is:
Aurora Biosciences  (US Phone: (858) 404-6600
http://www.aurorabio.com/

It works in flow (we do it), but I don't know if it is permeable into
bacteria.

CCF2 is a FRET dye so you collect both blue (cleaved) and green
(non-cleaved) fluorescence channels to measure activity.

Phil Marder


>From: "isabelle.hautefort" <isabelle.hautefort@bbsrc.ac.uk>
>To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
>Subject: B-lactamase assay in individual bacteria?
>Date: Tue, 6 Jun 2000 12:51:37 +0100
>
>
>dear all,
>I would like to detect B-lactamse activity in  individual bacterial cells
>so
>that I can control the presence of a (AmpR) plasmid and check if its copy
>number is rather similar in every cell. I know that there is a fluorescent
>substrate for b-lactamase (CCF2 like). Does anyone of you know whether it
>works
>in a flow cytometer (which channel?) and which supplier sales it? (not
>Molecular Probes, at least not yet).
>Thanks for your help.
>Regards
>Isabelle
>
>*********************************************************************
>Dr. Isabelle Hautefort
>Molecular Microbiology Section
>Institute of Food Research
>Research Park- Colney
>Norwich NR4 7UA- UK
>
>Tel +44(0)1603 255 391
>Fax +44(0)1603 458 414
>E-mail: isabelle.hautefort@bbsrc.ac.uk
>*********************************************************************
>

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