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Crosslinking Ab w/Antigen?


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#1 Bio Doc

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Posted 02 September 2009 - 04:52 AM

Greetings All,

I'm seeking some advice: I would like to permanently crosslink a monoclonal Ab I have against a known antigen. Keep in mind that this mAb will be biotinylated.

Here is the thing...would I be wise in just performing a quick UV crosslinking step in solution to help ensure they are permanently attached? The UV should not harm my biotin tag. For my experiment, I want to ensure the antigen:mAb remains permanently stable and I would like to avoid any unnecessary structural modifications. I want both the antibody and antigen to be in their most native states. Any insight would be greatly appreciated.

Any thoughts and/or suggestions would be greatly appreciated.

Cheers,

Bio Doc

#2 genehunter

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Posted 02 September 2009 - 04:58 AM

Did you mean that you will use Ag labeled with photoactivatable cross-linker plus UV treatment (prefered) or just UV? Pierce carries all kind of cross-linkers.

#3 Bio Doc

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Posted 02 September 2009 - 05:06 AM

Did you mean that you will use Ag labeled with photoactivatable cross-linker plus UV treatment (prefered) or just UV? Pierce carries all kind of cross-linkers.


At this point, I was considering just UV as I don't have the Ag labeled with any photoactivatable cross-linkers. I don't have tons of the mAb (as always tends to be the case), but what I do have is already biotinylated. At the least, the UV treatment shouldn't alter that.

Edited by Bio Doc, 02 September 2009 - 05:07 AM.





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