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" <[EMAIL PROTECTED]> 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:[EMAIL PROTECTED] On Behalf Of >> Gina Clayton >> Sent: Friday, June 27, 2008 7:37 PM >> To: CCP4BB@JISCMAIL.AC.UK >> 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: [EMAIL PROTECTED]
