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Co-IP suggestions w/o boiling at the end?

Co-IP suggestions w/o boiling at the end? - Immunoprecipitation Forum

Co-IP suggestions w/o boiling at the end? - IP or Immunoprecipitation Forum.


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Old 02-22-2009, 08:13 PM
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Default Co-IP suggestions w/o boiling at the end?



Hi everyone,

I will be doing a co-IP, but can't boil during the elution step, since these proteins aggregate when they are boiled, and thus show up at the wrong kDa size.

I was thinking of just rotating the beads in 5xSDS loading buffer @ 4degrees for half an hour, centrifuge, and load the supe onto my SDS-PAGE gel to be detected by western, but not sure if this will do the trick, or if anyone else have any other suggestions? I'm very new to this technique, any help will be of use!! Thanks!
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Old 03-11-2009, 06:54 PM
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Default Re: Co-IP suggestions w/o boiling at the end?

SDS works great.
As well as low pH or high salt.
google either low pH or high salt elution for typical buffer recipes.
Can even buy some from Pierce.
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