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DCED

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marthabrainard1

Contributor

Posts: 20

Joined: Wed Aug 25, 2021 2:10 pm

Post Mon Oct 18, 2021 4:22 pm

DCED

Hi, our lab is trying DCED instead of cisplatin as a viability stain. I read in the following paper that DCED was pre-conditioned with DMSO at 37C for 48 hours before freezing at -20C-. Could anyone give me details on these steps (how much DMSO you added and if you froze in DMSO, etc)? I'm assuming you can't freeze-thaw DCED so I didn't want to add too much DMSO to the powder.

Thank you

https://www.nature.com/articles/s41598- ... 91-2#Sec19
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Mon Oct 18, 2021 10:26 pm

Re: DCED

Hi Martha,

When this paper was still a preprint, Felix spoke at a Stanford CyTOF meeting. When I asked him in email after the meeting, he confirmed that:

"Yes, I treated it exactly as the cisplatin [in Fienberg et al], so resuspend to 100mM in DMSO and actually we also "pre-conditioned" both at 37C for 2 days (I took this from Sean but did not do direct comparisons if this is necessary). Afterwards, I froze it at -20C (it can be refrozen at least a few times) and I made pre-dilutions in PBS (1:200) which actually seemed to work great for at least 2 weeks but I did not thoroughly compare longer term storage."


So, make a 100mM stock of DCED in DMSO, condition it, and then freeze at -20C. Yes, I would also agree that repeated freezing and thawing probably isn't a good idea, so single-use aliquots are probably best. From the paper, the 500nM final concentration is a 200x dil of that 100mM stock; the staining was done in 1mL, so a single-use aliquot at 5uL of 100mM stock shouldn't be terrible to pipet....unless you do the entire 1g bottle at once (at MW 237.42 g/mol, 1g would make 42mL @ 100mM, if my math is right).


Mike
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marthabrainard1

Contributor

Posts: 20

Joined: Wed Aug 25, 2021 2:10 pm

Post Wed Oct 20, 2021 12:59 pm

Re: DCED

Thank you-this is super helpful!

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