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Stripped and recharged Ni-NTA beads but still getting impurities only with Ni-NTA beads

Sudheer Sangeetham via methods%40net.bio.net (by sudheer.pbm07 from gmail.com)
Fri Aug 4 05:18:08 EST 2017


Hi

*  Strip of nickel ions by washing with 10 bed volumes of 100 mM EDTA, pH
8.

* Wash resin with 10 bed volumes of de-ionized water.

* Charge resin with 2 bed volumes of 100 mM NiCl2 metal ion aqueous
solution.

* Wash resin with 10 bed volumes of de-ionized water to remove unbound
metal ions.

* Stored the resin in 30% etanol.

I loaded only recharged bead on the gel to check I could see nothing on the
sds-page or not, but I could see faint or visible proteins on sds page due
to protein and Ni didnt strip off completely.

please let me any solution for it.


Thank you


Regards
Sudheer


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