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Conjugating a primary metal antibody to a secondary antibody

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nameerabaig

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Posts: 7

Joined: Wed Nov 20, 2019 5:20 pm

Post Tue Sep 21, 2021 6:46 pm

Conjugating a primary metal antibody to a secondary antibody

Hello Forum,

Has anyone tried labeling metal conjugated antibody with a secondary antibody? I was wondering if a metal labeled primary antibody would be amenable to binding with a secondary antibody containing a fluorophore or would the polymer interfere with this type of conjugation with a secondary antibody? Thanks!
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roexx078

Participant

Posts: 8

Joined: Thu Feb 08, 2018 6:21 pm

Location: Nashville, TN, USA

Post Tue Sep 21, 2021 6:54 pm

Re: Conjugating a primary metal antibody to a secondary anti

We've done that when testing antibodies for imaging- we don't have a Hyperion on campus, so we do our MaxPAR conjugation, stain a test slide, and detect the antibody with a fluorescent secondary before shipping our IMC slides out for imaging. Of course, a negative result doesn't necessarily mean your antibody to target interaction is broken, the IgG may just not be detectable by the secondary. That said, we've had pretty good results with this approach, at least for QC of imaging reagents. We also stick the custom conjugate on positive comp beads and run on our suspension system, to make sure something in the tube has metal on it :-)

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