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Revisiting anti-GFP in suspension mass cytometry

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thinkCara

Participant

Posts: 5

Joined: Thu Sep 17, 2015 2:17 pm

Location: Bergen, Norway

Post Tue Sep 14, 2021 9:44 am

Revisiting anti-GFP in suspension mass cytometry

Hi All,

I'm looking for a good anti-eGFP clone that can be conjugated to a lanthanide for use with suspension mass cytometry. This topic was addressed in an older post without much success. Fluidigm no longer sells a conjugated anti-GFP. There are multiple clones out there.

Has anyone had success conjugating an anti-GFP antibody and getting good staining? Are there any publications with mass-tagged anti-GFP? I want to use it to help track GFP+ human cancer cells from a PDX mouse model.

Thanks for your help!

Best,
Cara
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cguidos

Contributor

Posts: 28

Joined: Tue Nov 18, 2014 3:10 am

Post Tue Sep 14, 2021 12:59 pm

Re: Revisiting anti-GFP in suspension mass cytometry

HI Cara

We found that the Fluidigm anti-GFP did not perform well, so purchased hybridoma for clone Sp2/0-Ag14 from the Developmental Studies Hybridoma Bank, University of Iowa (Cat #DSHB-GFP-4C9-f). We produced serum-free supernatant, purified IgG with Protein G Hi-trap column and metal tagged it with MAXPAR kit. It has worked well for us.

Hope this helps!

Cynthia
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Marjolyn

Participant

Posts: 9

Joined: Tue Sep 15, 2015 6:02 pm

Location: Leiden, The Netherlands

Post Tue Sep 14, 2021 1:36 pm

Re: Revisiting anti-GFP in suspension mass cytometry

Dear Cara,

This is a little bit more general but I have conjugated multiple antibodies that react to fluorophores. anti-BV605 and anti-PE-TxRed.
When there is an purified antibody available, it worked well for us.

Good luck.

Marjolijn

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