Immunoprecipitation native conditions

[nested]

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!

[diegogoy]

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

[nested]

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.