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Diluting antibodies in stabilization buffer

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AdeebR

Grand master

Posts: 169

Joined: Thu Mar 13, 2014 5:58 pm

Location: NYC

Post Wed Oct 01, 2014 3:06 pm

Diluting antibodies in stabilization buffer

Hi all,
Are there any concerns about over diluting antibodies in antibody stabilization buffer? I typically dilute to 0.2mg/ml until titration and then further dilute so that they can be used at ~1uL/test. In some cases, this second phase of dilution is quite significant (>1mL). Would this be expected to have any impact on long term stability?
Adeeb Rahman
Icahn School of Medicine at Mount Sinai, NYC
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mleipold

Guru

Posts: 5792

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Wed Oct 01, 2014 10:13 pm

Re: Diluting antibodies in stabilization buffer

Hi Adeeb,

There should not be any problem with that.

I've talked with Candor, and they recommend 100% storage buffer (ie, from your last spin filter step, use only storage buffer to bring back up to volume). Barring that, they recommend the more, the better.

Here at the HIMC, we often use 50% storage buffer: resuspend antibodies in 100% W buffer, spec by Nanodrop for protein concentration, then dilute with an equal volume of 100% stabilizer (for a net 50/50).

But I know of other people/labs who spin down again after Nanodrop, and resuspend in 100% stabilizer.


Mike
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MCOlivier

Contributor

Posts: 47

Joined: Mon Oct 05, 2015 9:48 am

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

Post Thu Jul 05, 2018 2:06 pm

Re: Diluting antibodies in stabilization buffer

Hi all,

I'm wondering which of the Candor AB Stabilization Solution is prefered (the PBS or the Tris one).
Any insight welcome ^^

Thx

Olivier
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mleipold

Guru

Posts: 5792

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Thu Jul 05, 2018 2:45 pm

Re: Diluting antibodies in stabilization buffer

Hi Olivier,

We use the PBS one. I haven't tried the Tris one.


Mike
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MCOlivier

Contributor

Posts: 47

Joined: Mon Oct 05, 2015 9:48 am

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

Post Thu Jul 05, 2018 2:47 pm

Re: Diluting antibodies in stabilization buffer

Thanks Mike !

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