Hi Monica=2C
If your protein is in the pellets I hope you are using denaturing condition=
s to solubilize it before purification over Ni-NTA.
solubilize the protein in urea or guanidine for one hour and spin down to g=
et a clear lysis solution before you proceed to further purification steps =
Ni-NTa or gel.
=20
cheers
wajahat
> Date: Fri=2C 4 Sep 2009 20:29:26 +0530
> From: monicamittal41 from yahoo.com> To: proteins from magpie.bio.indiana.edu> CC:=20
> Subject: [Protein-analysis] QUERY
>=20
> HI
> I'M FACING PROBLEM IN PURIFYING A protein which is a dna binding protein =
but its showing a kind of hydrophobic behaviour as its max part is going in=
to pellet after lysis.
> moreover its not responding to ni2+ nta column also and the lysis sol. is=
not so clear that i go for gel filteration as it may probably clog it
> so can u suggest any solution?
>=20
> monica
>=20
>=20
>=20
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