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Immunoprecipitation native conditions

Immunoprecipitation native conditions - Immunoprecipitation Forum

Immunoprecipitation native conditions - IP or Immunoprecipitation Forum.


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  #1  
Old 11-09-2009, 03:52 PM
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Default Immunoprecipitation native conditions



Hi,

I'm performing IP with Dynabeads Protein G. I'm trying to purify a protein with a V5 tag in it native state.

I've already tried elution in glicyne pH 2.7 (according to the recomended protocol) and nothing.

Anyone have a efficient protocol to elute proteins from dynabeads in its native state?

Thanks in advanced!
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Old 12-01-2010, 01:22 PM
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Default Re: Immunoprecipitation native conditions

Hey Nested
I have exactly the same problem, I need to elute my complex or my protein but in native state from the dynabeads. I did the elution with glycine 50 mM Glycine pH 2.8 at different times without any success, did you find a way to do perform the elution. Thanks in advance for your help
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Old 01-04-2011, 03:53 PM
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Default Re: Immunoprecipitation native conditions

Hi,

Unfortunately I didn’t find a way to elute the protein from the beads in it native state. I read somewhere that you can´t perform an elution with glycine if you crosslink the antibody . So I tried to do the elution without the crosslinker but nothing...

I also tried it with glycine pH 13, and I could get a very very weak band in my blot but only 1 or 2 times... So if by chance you find a reproducible way to do it please let me know.
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