DRAQ5: may be an alternative for Hoechst 33342 if I have only 488nm argon laser?

Derek Davies daviesd2 at cancer.org.uk
Thu Mar 7 10:04:08 EST 2002


Hello Pal,

Yes, we have used DRAQ5 here as a vital DNA dye. You are right that on
excitation by a 488 Argon laser it emits in the far red so on a FACScan
or single laser FACS Calibur it will be detected in FL3. I have attached
a small graphic file to show Jurkat cells unstained and stained with 5uM
DRAQ5 for 2mins. I have run these on a Calibur with log amplification
for both FL2 and FL3 to show that no compensation is needed to remove
DRAQ5 signal from the FL2 channel. Obviously if you were using it for
quantitative DNA assessment, you would use linear amplification. There
would still be some compensation needed to remove bright PE signals from
FL3.

We have used DRAQ in combination with GFP and Pyronin Y so it can be
done!

Derek


On Wed, 6 Mar 2002, Jáksó Pál wrote:
> I read a post from Derek Davis in which DRAQ5  a far red emitting DNA
> stain from Biostatus was recommended for DNA staining for flow cytometry
> without permeabilization of living cells. I would like to ask about your
> experiences with this new DNA dye.
> I intend to measure surface immunostainings in conjuctions with DNA
> content using 488 nm argon laser. At the Biostaus website is stated that
> using together with FITC compensation is not necessary and DRAQ5 can be
> also used with PE.

************************************************************************
Derek Davies                       Voice: (44) 020 7269 3394
FACS Laboratory,                   FAX: (44) 020 7269 3100
Cancer Research UK,                e_mail:derek.davies at cancer.org.uk
London Research Institute, 	   mobile: 07790 604112
44 Lincolns Inn Fields,
London, UK.

Web Page: http://sci.cancerresearchuk.org/axp/facs/davies/index.html

In tenebris lux 							 
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