Re: H2DCFDA

From: Arnold Pizzey (a.pizzey@ucl.ac.uk)
Date: Mon Apr 03 2000 - 03:33:55 EST


At 12:05 31/03/00 +0100, you wrote:
>
>Hi!
>
>We are interested in measuring cellular ROS production by the use of
>H2DCFDA in flowcytometry. Company instructions indicate that the dissolved
>reagent has a short stability (few days). The product is provided in 100mg
>size package. Thus, on one hand it is too difficult  (and overall too
>harmful) to weight small quantities, and, on the other, if it is necessary
>to dissolve the whole amount for each test, the cost of experiments will be
>too high.
>We would like to know your experience  about the storage conditions of
>H2DCFDA, and if anyone has tested the stability of the dissolved reagent.
>Thanks .
>
>Bye
>
>Angela & Anna
>
>Dr. Walter Giaretti
>Laboratory of Biophysics and Cytometry
>National  Cancer Institute (IST)
>Largo Rosanna Benzi, n. 10
>16132 Genoa, Italy
>
>Tel. +39-10-5600969
>Fax  +39-10-5600711
>E-mail  giaretti@hp380.ist.unige.it

Greetings Angela & Anna,

I have used the above reagent (molecular probes D-399) for the measurement
of granulocyte ROS in a whole blood assay. It has been my experience that
the reagent is unstable once dissolved in DMSO; typically we would make it
up on the day of the experiment and discard any unused.

We have been using a 100mg bottle of the above on and off for a few years
now- it never seems to run out and at 130K Lira it couldn't be much
cheaper. Also as far as I can determine, there are no particular safety
issues when handling this substance.

Best regards,

Arnold


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	Arnold Richard Pizzey
	Department of Haematology
	Royal Free and University College London Medical School
	98 Chenies Mews
	London WC1E 6HX
	U.K

	voice:	+44 0207-209-6234
	Fax:	+44 0207-209-6222
	email:	a.pizzey@ucl.ac.uk
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