Nah, I have one that can handle 2.5 liters.
Peter J. Miller
Collins Laboratory
Department of Biochemistry and Biophysics
University of North Carolina at Chapel Hill
919-966-9410
On 6/30/08 11:33 AM, "Gina Clayton" <[log in to unmask]> wrote:
> Hi there
>
> I actually just consulted, about your question, with one of the
> longer term members of the department about this. And we came to the
> conclusion that 4- 500ml was probably the maximum size.
>
> Gina
>
> On Jun 30, 2008, at 11:15 AM, Radisky, Evette S., Ph.D. wrote:
>
>> Another question re: Amicon stirred cells...
>>
>> I also seem to recall seeing 1L size stirred cells in older labs of my
>> youth. My current lab has acquired one of 400 mL, but looking to
>> purchase a bigger one, I can't find any. Any ideas about where we
>> might
>> find one?
>>
>>
>> Evette S. Radisky, Ph.D.
>> Assistant Professor and Associate Consultant II
>> Mayo Clinic Cancer Center
>> Griffin Cancer Research Building, Rm 310
>> 4500 San Pablo Road
>> Jacksonville, FL 32224
>> (904) 953-6372 (office)
>> (904) 953-0046 (lab)
>>
>> -----Original Message-----
>> From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of
>> Gina Clayton
>> Sent: Friday, June 27, 2008 7:37 PM
>> To: [log in to unmask]
>> Subject: Re: [ccp4bb] Concentrating protein
>>
>> Hi there
>>
>> I quite like the Amicon stirred ultra concentration cell systems. You
>> can put large volumes in, maximum 1 litre size, I think. As well you
>> can attach an inert gas such as Argon or Nitrogen, for the gaseous
>> pressure, this reduces oxidation of your sample while it
>> concentrates. My experience has been that, depending on the filter,
>> the filters are very resistant to various salts even GuHCl, and you
>> get
>> good recovery. I used to concentrate large volumes of protein down to
>> say 50-25ml then switch to the same system, in a much smaller cell
>> i.e.
>> 10ml, to get down to 1-2ml. And they are fairly fast too.
>>
>> I get the impression, perhaps incorrectly, they are not as
>> fashionable
>> as they used to be, but perhaps "older labs" tend to have them
>> milling
>> about somewhere in the back of a cupboard. So most likely you would
>> only have to buy membranes -PM or YM it think depending on you
>> sample.
>>
>> Hope that helps
>>
>> Gina
>>
>>
>>
>>
>> On Jun 27, 2008, at 9:19 AM, Roger Rowlett wrote:
>>
>>> Guenter Fritz wrote:
>>>> A mild and quick method is to use dry Sephadex G-25. The material
>>>> will swell and take up all the liquid except molecules larger than
>>>> ca.
>>>> 5 kDa.
>>>>
>>>>
>>>>> Dear All,
>>>>>
>>>>> we have GCSF protein produced in inclusion bodies. we solubilise it
>>>>> refold it and then concentrate it using proflux system. still the
>>>>> concentration of the protein we get is less and volume is more for
>>>>> us to load in Ion exchange chromatography. is there any simple
>>>>> technique that can be performed in lab without using any hi-fi
>>>>> instrument to concentrate the protein in small volume of buffer.
>>>>> the
>>
>>>>> protein we obtain is about
>>>>> 0.7
>>>>> mg/ml and we get 450 ml solution. our column is 110ml lab scale and
>>>>> we have to work in that only. i have heard of NH4SO4 precipitation.
>>>>> but it
>>>>> requires protein conc more than 1 mg/ml.
>>>>>
>>>>> kindly help me to progress in my experiment.
>>>>>
>>>>>
>>> One of the beauties of ion-exchange chromatography is that it is an
>>> excellent concentration step as well as a purification methodology.
>>> It may take less time and involve less protein loss to pass all the
>>> solution through the IEX column and bind the protein, assuming you
>>> have the protein in a low ionic strength buffer at the appropriate
>>> pH.
>>
>>> Elution in a smaller volume can be accomplished by increasing the
>>> NaCl
>>
>>> concentration to an appropriate level. In the bad old days before
>>> bacterial overexpression, we used to to this routinely to concentrate
>>> a liter or more of protein extract to 50-100 mL after elution from a
>>> small, high-capacity IEX column.
>>>
>>> Cheers,
>>>
>>>
>>> --
>>> ---------------------------------------------------------------------
>>> -
>>
>>> --
>>> Roger S. Rowlett
>>> Professor
>>> Colgate University Presidential Scholar
>>> Department of Chemistry
>>> Colgate University
>>> 13 Oak Drive
>>> Hamilton, NY 13346
>>>
>>> tel: (315)-228-7245
>>> ofc: (315)-228-7395
>>> fax: (315)-228-7935
>>> email: [log in to unmask]
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