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Immunoprecipitation overnite or for several hours?

Immunoprecipitation overnite or for several hours? - Protein Science

Immunoprecipitation overnite or for several hours? - Post Questions and Discuss Protein Structure and Function, Protein Localization etc.. Please do not post Proteomic questions here, there is a Proteomic Forum below.


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Old 12-05-2006, 07:52 AM
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Cool Immunoprecipitation overnite or for several hours?



Hey guys,
should I immunoprecipitate overnite or for several hours?

thank u
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Old 12-19-2006, 11:40 PM
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Default Re: Immunoprecipitation overnite or for several hours?

It depends on several variables, quality of the antibody, affinity and specificity, stringency of the incubation buffer, abundance of your protein, to name a few.
If this is the first time you use this method I would recommend overnight incubation at 4C, and if the signal is strong you could try cutting down the incubation times.
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Old 07-15-2007, 09:46 AM
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Default Re: Immunoprecipitation overnite or for several hours?

generally, IP for 4 hours is good enough.
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Old 07-17-2007, 08:08 PM
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Default Re: Immunoprecipitation overnite or for several hours?

ON is safe if your antibody is specific and degradation is controlled via 4C or Proteinase inhibition. If your prep is clean and not a biological fluid you should be fine with ON. Biological fluids have endogenous antibodies that will bind to protein A/G and co elute with your eluant--learned this the hard way.
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