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Recombinant Protein precipitate out under 20 mM KPB/100 mM NaCl ???

Recombinant Protein precipitate out under 20 mM KPB/100 mM NaCl ??? - Protein Forum

Recombinant Protein precipitate out under 20 mM KPB/100 mM NaCl ??? - Protein Forum


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  #1  
Old 02-15-2008, 12:21 AM
Nico
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Default Recombinant Protein precipitate out under 20 mM KPB/100 mM NaCl ???



Hi,

I am changing buffer for my protein, which was purified from Ni-NTA column.
Under 20mM KPB/500mM NaCl, it was good. But when change to 20 mM KPB/100 mM NaCl,
the protein precipitate out. What should I do? Change to Tris-HCl Buffer or.....

Please help me out,

Many thanks,

Nico


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Old 02-18-2008, 09:37 PM
Dr Engelbert Buxbaum
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Default Recombinant Protein precipitate out under 20 mM KPB/100 mM NaCl ???

Am 14.02.2008, 20:21 Uhr, schrieb Nico <[Only registered users see links. ]>:



Looks like your protein is an albumin, with elongated shape and asymetric
charge distribution. At low ionic strength they form head-to-tail
aggregates, which precipitate. You can either keep the ionic strength high
or change the pH and hence the charges on the protein.
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  #3  
Old 02-20-2008, 02:14 PM
incrediblyhigh@hotmail.com
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Default Recombinant Protein precipitate out under 20 mM KPB/100 mM NaCl???

On Feb 18, 9:37*pm, "Dr Engelbert Buxbaum"
<[Only registered users see links. ]> wrote:

I agree with Dr Buxbaum. Try increasing the NaCl concentration (you
could try between 200-500mM if you really need to drop the
concentration), clearly the main reason for the precipitation.
Changing the buffer/pH will probably not help much...
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kpb or 100 , nacl , precipitate , protein , recombinant


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