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Freezing Antibodies

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atsponaugle

Participant

Posts: 4

Joined: Thu Jul 01, 2021 5:31 pm

Post Tue Jul 06, 2021 1:47 pm

Freezing Antibodies

Hi,

I am doing a longitudinal study and would like to make a large batch of 1) surface and 2) intracellular antibodies that I can use for the entire study. Two questions:
1. How do you recommend freezing surface antibody cocktails (buffer, volume, etc)?
2. Is it possible to freeze intracellular antibodies?

Thanks,
Alexis
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mleipold

Guru

Posts: 5792

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Tue Jul 06, 2021 9:10 pm

Re: Freezing Antibodies

HI Alexis,

You can find info about how to freeze antibody cocktails in 2019-Schulz et al-Cytometry A here: https://doi.org/10.1002/cyto.a.23781

There isn't a difference in freezing intracellular vs surface antibodies. But it's worth doing a test to make sure that all your antibodies survive at -80C (so far all of ours have, but it's worth a week to find out before you spend a lot of time and money).

Volume is up to you. Single-reaction aliquots give you maximum flexibility for variable sample number, but may mean you're pipetting 100+ tubes to freeze. If you have a standard number of samples per plate/batch, you might consider making one tube per plate (at the appropriate volume).


Mike
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atsponaugle

Participant

Posts: 4

Joined: Thu Jul 01, 2021 5:31 pm

Post Wed Jul 07, 2021 11:22 am

Re: Freezing Antibodies

Hi Mike,

Thanks so much for the reply. What buffer do you typically freeze both surface and intracellular antibodies at?

Alexis
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mleipold

Guru

Posts: 5792

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Wed Jul 07, 2021 12:36 pm

Re: Freezing Antibodies

Hi Alexis,

I've usually used CyFACS (PBS+EDTA+BSA). I think others have used Fluidigm's cell staining buffer, but maybe people can chime in about that.


Mike
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atsponaugle

Participant

Posts: 4

Joined: Thu Jul 01, 2021 5:31 pm

Post Wed Jul 07, 2021 12:57 pm

Re: Freezing Antibodies

Hi Mike,

Do you usually keep intracellular markers in CyFACS as well?

Thanks,
Alexis

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