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Understanding immunoprecipitation - where do the beads go?

beads immunoprecipitation IP agarose antibody

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#1 DrZeus

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Posted 07 May 2013 - 09:38 AM

Hi, I know this may be a very mundane question but I cant seem to find it anywhere. I'm just wondering where the beads go after you run an IP sample on SDS-PAGE? As in the protein/antibody complex are loaded as I have blotted against the same antibody used in the IP and I can see the heavy and light chain bands, but I was wondering where the beads go? Do they get removed in the clearing spins, or when i add SDS loading buffer?
Thanks very much

#2 GNANA

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Posted 07 May 2013 - 11:21 AM

The final step of IP is to add the loading buffer onto beads-Ag-Ab complex, you ll then denature it by boiling, so here the antibodies and the antigen will get eluted from the beads, after denaturation beads are spinned down and loaded only the supernatant, which contains antigen and fragmented antibody.
I would prefer being perfectionist rather than a passionist in Research.

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#3 @bhijit

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Posted 04 June 2013 - 10:16 PM

Is there any way to get only the antigen from beads-Ag-Ab complex ???





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