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Getting monoisotopic cisplatin Pt195 and 196

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It is fine to promote your company's reagents. Just make sure they are relevant to CyTOF, and do so in moderation and style :-)
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kanthsmiles

Participant

Posts: 7

Joined: Sun May 11, 2014 2:41 pm

Post Thu Aug 08, 2019 9:13 pm

Re: Getting monoisotopic cisplatin Pt195 and 196

Thanks a ton Mike! This definitely gives a better understanding.
Regards
Kanth
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hartmannfj

Participant

Posts: 7

Joined: Fri Oct 31, 2014 3:51 pm

Post Mon Aug 12, 2019 5:43 pm

Re: Getting monoisotopic cisplatin Pt195 and 196

Sorry I just saw this now and thanks a lot to Mike, I couldn't have explained it better. I just wanted to say that the preconditioning step is something that we do for all cisplatin arriving in the lab (also the one for live/dead discrimination) and was established by Sean I believe. Personally, I did not compare labeling efficacy with and without preconditioning but all our labelings are done with preconditioned cisplatin so it definitely works well.

Best, Felix
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JamesW

Contributor

Posts: 26

Joined: Tue Nov 21, 2017 6:59 am

Post Fri Aug 16, 2019 6:08 am

Re: Getting monoisotopic cisplatin Pt195 and 196

Hello all,

Just to chime in on this one previously I did 48 hours of preconditioning on DCED-palladium based on Felix's paper but this time I skipped it as my cisplatin got stuck in a shipping warehouse for several days and was pretty warm on arrival. I just aliquoted and put it in the Freezer.

At least my understanding of the "Say no to DMSO" paper linked in Mikes comment is that the observed loss of cytotoxicity must be pretty rapid as I think the authors just dissolve the cisplatin in DMSO right before use. Presumably, the cytotoxicity drops further with the suggested 48 hours of preconditioning but I don't have any information on how much.

Jahangir, I did just follow the Mei et al. protocol for the antibody labelling, it seems to have worked well.

Best,
James
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henkmei

Participant

Posts: 16

Joined: Wed Jul 30, 2014 9:44 pm

Post Fri Aug 16, 2019 7:05 am

Re: Getting monoisotopic cisplatin Pt195 and 196

Hi all,

I am really glad to see that there is much interest in the platinum conjugates.

To the end of this discussion,I simply want ot state that we did not try preconditioning the cisplatin solutions we use for dead cell labeling and antibody conjugations. We still use the protocol as published in Cytometry A https://onlinelibrary.wiley.com/doi/ful ... to.a.22778 with consistently good results, so we did not have a reason to put efforts into further optimization.

Best
Henrik
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MCOlivier

Contributor

Posts: 32

Joined: Mon Oct 05, 2015 9:48 am

Location: European Genomic Institute for Diabetes, Universitity of Lille, Institut Pasteur de Lille, France

Post Mon Aug 19, 2019 7:32 am

Re: Getting monoisotopic cisplatin Pt195 and 196

Hi Cisplatin-geeks ^^

I was on holidays so I answer quiet late... Sorry.

I did follow the protocol from the Mei et al, Cytometry A, 2015 paper, without any pretreatment.
For storage, basically, what I was told by Petr Vasiliev from Buyisotopes was that Cisplatin is very sensitive to heat (and light !), and that freezing/thawing should be totally avoided !!! Consequently, I did aliquots like you Jahangir, except that I kept the Cisplatins at 10mM and made aliquots of 2µL, as for 1 AB conjugation reaction, and store them in -20°C. Quiet a long task as we got 1.5 to 2 ml :)

To be continued with the 194/195 Cisplatins ;-)

Olivier
(Lots of discussions to read after 3 weeks !)
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