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L-Buffer alternative

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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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mds4z

Participant

Posts: 9

Joined: Tue Dec 03, 2013 6:23 pm

Post Fri May 17, 2024 1:23 pm

L-Buffer alternative

Hello

We are in need of extra L-buffer. Were told can't purchase this individual, only with kits from Biotools. Is there an alternative source, or recipe to make your own?

Thanks
Mike
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DMcDonald

Contributor

Posts: 23

Joined: Thu Nov 26, 2015 10:14 am

Post Fri May 17, 2024 3:15 pm

Re: L-Buffer alternative

Hi Mike,

I've a sneaky suspicion it's just 150 mM sodium phosphate pH7.2. I could be wrong but it's from this paper which describes a near identical conjugation and I've always assumed the original DVS kit spawned out of: https://pubmed.ncbi.nlm.nih.gov/20939532/

I've never tried the switch though.

David
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ezunder

Participant

Posts: 5

Joined: Tue Oct 03, 2017 2:10 pm

Post Fri May 17, 2024 3:57 pm

Re: L-Buffer alternative

Hi,

David- I think what you put here is the R-buffer, not the L-buffer, right? The paper you cited is great! They use 150 mM sodium phosphate buffer at pH 7.2 as an R-buffer (reducing) for the TCEP step. The L-buffer (loading) is 20 mM ammonium acetate at pH 5.2. This is used for DTPA/Lanthanide incubation.

If I recall correctly, we never had to pH the ammonium acetate because it was around pH 5-6 after dissolving in water. The low pH helps prevent metal oxidation I think.

I vaguely remember hearing that DVS/Standard Biotools switched away from using Ammonium Acetate as their L-buffer, but this may just be hearsay :)

We always used L-buffer for metal storage as well as DTPA loading. I bet that Ammonium Acetate is still good for DTPA loading, but maybe they found something better for long term storage?

Mike- if you decide to use Ammonium Acetate for L-buffer, I would buy the purest you can find from Sigma-Aldrich and definitely run a small aliquot in liquid mode to make sure there's no contaminating metals before using it for antibody conjugations! :)

Best,
Eli
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DMcDonald

Contributor

Posts: 23

Joined: Thu Nov 26, 2015 10:14 am

Post Fri May 17, 2024 4:06 pm

Re: L-Buffer alternative

Hi Eli, Yes you're right I've got them switched!
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CRStevens

Master

Posts: 61

Joined: Thu Jul 17, 2014 5:07 pm

Post Fri May 17, 2024 4:19 pm

Re: L-Buffer alternative

Mike,

My first question to you is do you believe L-buffer has an expiration date. I have some vintage L-buffer. I'm not sure if it is like wine that gets better with age but let me know how picky you are.

-Chad

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