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Alternative sources for isotopes

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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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Kjwaller

Participant

Posts: 11

Joined: Sat Aug 27, 2016 6:56 am

Post Tue Feb 13, 2018 1:13 pm

Re: Alternative sources for isotopes

Hi,

I just wanted to update on my CD3 89Y conjugation using the yytrium from sigma. I haven't got the data together but the conjugation went really well, I am running it through cytof as we speak. It's actually much better than the conjugation I was given from fluidigm and about a tenth of the price, so thanks very much for giving me the advice and confidence to give it a go!

Great forum!

Kate
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jlederer

Participant

Posts: 8

Joined: Thu Feb 08, 2018 2:42 pm

Post Tue Feb 13, 2018 5:10 pm

Re: Alternative sources for isotopes

We can label with 89Y, but it is not stable. Be careful and check for stability in your labeled CD3 Ab. Good luck. Jim
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jeccles

Participant

Posts: 5

Joined: Thu Jul 14, 2016 4:07 am

Post Tue Feb 13, 2018 5:19 pm

Re: Alternative sources for isotopes

Hi Kate-
Can you explain how you manage to spend 1/10 of the price? Don't you still need to buy a fluidigm conjugation kit to get polymer? Or is there some cheaper way to purchase polymer alone? Thanks for working on this!
- jake
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GregBehbehani

Master

Posts: 80

Joined: Tue Apr 12, 2016 10:17 pm

Location: The Ohio State University, Columbus, Ohio

Post Tue Feb 13, 2018 6:05 pm

Re: Alternative sources for isotopes

Hi Kate and Jim,

First off, Kate, thanks for sharing your results with the group. It's good to know things worked well for you. We're all happy to help.

Jim, I'm curious about your stability issues with 89Y. We have used an anti-CD235 antibody on 89Y for a little over a year, without any appreciable loss of function. I'd love to learn more about the problems you had.

best,

Greg
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jlederer

Participant

Posts: 8

Joined: Thu Feb 08, 2018 2:42 pm

Post Tue Feb 13, 2018 7:01 pm

Re: Alternative sources for isotopes

Hi Greg. Good to hear that you have had stable 89Y labeling. We tried it with several dfferent antibodies. We always saw a steady loss of retention of 89Y in the X8 polymer. I was thinking that maybe it is the way that we dissolve and store the metal. Can you share with the group how you prepare 89Y for your conjugations? That might be the difference. We also gave up on 139La. Best regards, Jim
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GregBehbehani

Master

Posts: 80

Joined: Tue Apr 12, 2016 10:17 pm

Location: The Ohio State University, Columbus, Ohio

Post Tue Feb 13, 2018 7:10 pm

Re: Alternative sources for isotopes

Hi Jim,

Thanks for the information. I'll double-check the details from my technicians, but we usually just dissolve the chloride we get from Sigma in L-buffer at 1M (or the highest concentration we can achieve) we then dilute further to make a working stock. Occasionally, we have to add a bit of acid (usually HCL or HNO3 to get the metal to dissolve), but I don't think we needed to do this for 89Y. We keep our metals in the refrigerator (a hold-over from what we did in the Nolan Lab).

I believe the L-Buffer is important, since you want to be sure the metal solution stays acidic, but I otherwise can't think of a reason that things wouldn't have worked for you.

We'll go back and re-test our 89Y and 139La antibodies to see if we're starting to see any subtle loss of signal, but they definitely still work as of a few weeks ago.

I'll post some follow-up once we finish our tests.

best,

Greg
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nkhanbham

Master

Posts: 53

Joined: Wed Feb 25, 2015 3:03 pm

Post Wed Feb 14, 2018 3:38 pm

Re: Alternative sources for isotopes

This is a very helpful thread.
I am going to try 89Y conjugation using the Sigma metal. Where are people buying their polymer from?
Thanks to all contributors.
Bw,
Naeem
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jeccles

Participant

Posts: 5

Joined: Thu Jul 14, 2016 4:07 am

Post Thu Feb 15, 2018 5:14 pm

Re: Alternative sources for isotopes

Exactly: "Where are people buying their polymer from?" This discussion seems to take this reagent for granted. Do you all have special access? If so, please be honest and acknowledge-
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mleipold

Guru

Posts: 2086

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Thu Feb 15, 2018 5:25 pm

Re: Alternative sources for isotopes

The HIMC buys our polymer as part of MAXPAR kits; there is no other commercial supplier as far as I'm aware, and to my knowledge Fluidigm still does not sell polymer-only (you have to get it with the metals and buffers in a MAXPAR kit).

Due to the number of in-house/custom conjugations we do, we go through enough polymer that we basically buy exclusively the 40rxn kits, either as a one-metal or as part of the Multimetal kits (#201300).

While the up-front cost of a 40rxn kit is high, it's almost a 50% savings on a per-reaction basis relative to the 4rxn kit. The Multimetal per-reaction cost is about halfway between the 40rxn and the 4rxn (as one-metal orders), but does give you the versatility of ordering up to 10 metals of 4 reactions each....slightly reducing the barrier for entry, or making it easier to restock certain metals that you happen to go through more quickly.
Last edited by mleipold on Fri Feb 16, 2018 12:20 am, edited 1 time in total.
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jlederer

Participant

Posts: 8

Joined: Thu Feb 08, 2018 2:42 pm

Post Thu Feb 15, 2018 11:00 pm

Re: Alternative sources for isotopes

Mike is correct. Fluidigm will not sell polymer alone. We also buy the 40 metal kits because we do a lot of conjugations for our CyTOF antibody resource and for other investigators. It is a significant saving to order the multimetal kits. Obviously, you can indicate which metals you want. We keep an inventory and just order kits with the metals that we need, sometimes we order the same metals in multiples. Jim
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