Well, DRAQ5 seems to be the flavour of the week - I speak as a relatively new user of the dye and I am sure Paul Smith or Terry Hoy at Cardiff are more qualified to answer but that has never stopped me before, so... On Mon, 11 Mar 2002, simon Monard wrote: > Do you know how far in the red DRAQ5 emits, can you use it on a sorter > as well as PE/Cy5 and collect the DRAQ5 above 710nm or something? As far as I know the max emission of DRAQ5 is about 700nm but it extends up to at least 800nm. I just did a quick and dirty experiment using PE-CY5 and DRAQ5 with 488nm excitation - collecting PE-Cy5 through a 670/20 filter and DRAQ5 through a 700LP filter using a 710DCLP to separate the signals, there was a bit of overlap of DRAQ5 into the PE-CY5 channel which needed about 5% compesation, none needed the other way but it was a relatively low PE-CY5 signal so maybe with a brigher one it might be necessary. This was the only filter combination readily to hand but it did look encouraging and I am sure with a little tweaking of the filter set-up it can be done (there may be a trade off of a slightly fatter DNA CV though). I'll get back to you when I have explored it a bit more. How are the chickens by the way? On Mon, 11 Mar 2002, Mostowski, Howard S. wrote: > DRAQ5 sounds as the next best thing since sliced bread; therefore I > have several questions. > > 1. From whom can it be purchased from [USA}? Alexis Biochemicals (as in the UK) 6181 Cornerstone Ct. E., Ste 103 San Diego, CA USA 92121 Tel: 800-900-0065 Fax: 800-900-9224 Email: alexis-usa@alexis-corp.com Web: www.alexis-corp.com > 2. Can it also be used as a live/dead discriminator, since > it couples with DNA? The staining of live cell DNA is very rapid, generally within 2-5mins, so this rather means it hasnt proved that useful in our hands as a live-dead discriminator. > 3. Are there any dangers in handling namely carcinogenic? It binds DNA so potentially harm,ful, take the usual precautions as when handling these types of dyes. More information (including references) at: http://www.biostatus.co.uk/draq5.html Don Rosson asked about using the dye on large scale. I cant say that I have done that but even in analytical experiments, we have seen that the profiles are quite dye and cell concentration dependent. As a guide, I get good HL60 profiles when using 5uM DRAQ to 10e6 cells. Changing either dye or cell concentrations by a factor of 2 results in a worse CV so I suspect that you would need to be quite careful as the optimal range may be quite narrow. Derek ************************************************************************ Derek Davies Voice: (44) 020 7269 3394 FACS Laboratory, FAX: (44) 020 7269 3100 Cancer Research UK, e_mail:derek.davies@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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