[Protein-analysis] Re: problem of concentrating protein
sandy_zhu_2006 from hotmail.com
(by sandy_zhu_2006 from hotmail.com)
Sat Jan 19 07:29:04 EST 2008
On 1=D4=C217=C8=D5, =C9=CF=CE=E73=CA=B118=B7=D6, "Dr Engelbert Buxbaum"
<engelbert_buxb... from hotmail.com> wrote:
> Am 10.01.2008, 01:01 Uhr, schrieb <sandy_zhu_2... from hotmail.com>:
> > The protein I am purifying is a kind of zinc finger protein which
> > has 81 amino acid and its theoretical pI is 9.19.But after purified
> > with Hitrap Heparin column the target protein were still stable,then
> > when I use the binding buffer(20mM Tris,10mM Mer,100mM NaCl and 1mM
> > ZnCl2,pH 7.0)to dialyze in order to remove the 2M NaCl which were used
> > in eluting the protein from the Heparin column, I found that there are
> > many sediment in it.
> > How to solve this problem?Is there any buffer which is more siutable
> > than Tris? Since at pH7.0 Tris is not as efficient as it was.
> > Plus I tried phosphate buffer,but it will interact with the ion
> > zinc (within zinc finger protein) and produce sendiment too.
> What is the composition of your elution buffer? Does it contain Zn? Does =
> the precipitation also occur when the binding buffer does not contain Zn?
My elution buffer do have Zn(1mM ZnCl2) in it.How to improve its
is the mayor concern right now.
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