Hi,
Could you not carry out Ammonium Sulfate precipitation to concentrate the
protein? I never did it myself so I have no idea if it's practical in this
case. Maybe it's still worth mentioning, though.
Tim
--
Tim Gruene
Institut fuer anorganische Chemie
Tammannstr. 4
D-37077 Goettingen
GPG Key ID = A46BEE1A
On Fri, 4 Sep 2009, Radovan Spurny wrote:
> Stirred cell from Milipore is very useful for concentrating big volumes of protein solution. There are cells from 3 to 400 mL. Ultrafiltration is performed through the membrane with high pressure by connecting to inert gas (nitrogen). It is very fast and comfortable technique. Here are more information: http://www.millipore.com/catalogue/module/C3259
>
> Sincerely,
>
> Radovan Spurny, PhD
> Laboratory of Structural Neurobiology
> Division of Pharmacology
> Campus Gasthuisberg, ON1
> Herestraat 49, PB 601
> B-3000 Leuven
> Belgium
> e-mail: [log in to unmask]
> ________________________________________
> From: CCP4 bulletin board [[log in to unmask]] On Behalf Of Yingyun Liu [[log in to unmask]]
> Sent: Thursday, September 03, 2009 16:48
> To: [log in to unmask]
> Subject: Re: [ccp4bb] Devices Suitable to Concentrate Protein Solutions in Liter Scale
>
> An anion exchange column with a large enough bed volume might be used to
> concentrate the protein. You probably need to change the pH of the
> solution so that the protein sticks to the column.
>
> On Thu, 2009-09-03 at 17:05 +0800, Xuan Yang wrote:
>> Dear Dr Ku,
>>
>> It was a wonderful idea! However, the refolding buffer contained 500mM
>> L-Arg and 1mM EDTA.
>>
>> And I want to try other affinity columns, just don't know what resin
>> would be appropriate.
>>
>> Sincerely,
>>
>> Xuan Yang
>>
>>
>> 2009/9/3 Shao-Yang Ku <[log in to unmask]>
>> Can you put a 6His-tag on your protein and add some Ni-NTA
>> beads to your solution to capture (and concentrate) the
>> properly folded molecules?
>>
>>
>>
>> Quoting "Xuan Yang" <[log in to unmask]>:
>>
>> Dear All,
>>
>> I am working on protein refolding via dialysis in
>> large volumn
>> (typically 2~4 litters). It was problematic when I
>> wanted to concentrate the
>> solution to at least less than 500ml. If you know any
>> device appropriate for
>> such task, please help me out:)
>>
>> Thanks in advance!
>>
>> Sincerely,
>>
>> Xuan Yang
>>
>>
>>
>>
>
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