Hi,

I suggest you to use Centricon Plus-70 Centrifugal Filter for your large volume of protein concentration. The tube volume is 70 ml and its the maximum volume and you can reuse the coloum for the same protein extract.

Thank you Kannan for your valuable suggestion. Can you tell in brief how we can centricon 70 and how many times we can reuse it.

Hi,

You can add your protein fraction into the centricon tube which has the filter inside the falcon and use the centrifuge and discard flow throw and your protein will remains in the filter collect the final fraction and repeat the same until you finish with your protein solution.

You can find the centricon tube link here.

https://www.sigmaaldrich.com/catalog/product/mm/ufc710008?lang=de®ion=DE.

You can store the colum with 10% EtOH wash with PBS 3 times and reuse the colum.

Thank you Kannan for your inputs.

https://www.sigmaaldrich.com/labware/labware-products.html?TablePage=21942948

Have you considered using an Amicon stirred cell (http://www.merckmillipore.com/DE/de/life-science-research/protein-sample-preparation/protein-concentration/large-volumes-samples/amicon-stirred-cells/tYyb.qB.CPEAAAFBxwtlvyxv,nav)? Works for both concentration and buffer exchange.

I do agree with Engelbert Buxbaum. Amicon is a nice system for the volume you're dealing with. Anyway, you must be ready to loose a big part of your protein sample during concentration steps (doesn't matter If it will be to dialysis bags or Amicon systems).

A few options below:

https://www.nist.gov/sites/default/files/documents/ncnr/StirredUltrafiltrationCell_Millipore.pdf

https://www.coleparmer.com/c/stirred-cells

http://www.advantecmfs.com/filtration/accessories/stirredcells.php

https://www.fishersci.com/us/en/products/I9C8KNMP/stirred-cells.html

http://www.interchim.fr/ft/B/BB131a.pdf

https://laboratory.pall.com/content/dam/pall/laboratory/literature-library/non-gated/pdf_1489182949676.pdf

http://www.deneysel.com.tr/UserFiles/File/Pdf/Cihazlar/Cevre-ve-Su-Analizleri/Yuksek-Basincli-Membran-Filtrasyon-Sistemi.pdf

http://www.amsmembrane.com/en/products/stirred_cell_hpc-197_eng

Hello Kumar;

If you will be dealing with this kind of separations on a regular basis, I suggest that you consider a larger scale cross-flow ultrafiltration system such as Pellicon 2 Mini cassette from Merck- Millipore:

http://www.merckmillipore.com/MX/es/product/Pellicon-Mini-Cassette-Holder,MM_NF-C614#

With such filtration system you can start with 0.1m^2 cassettes (up to 0.3 m^2) for protein separation from about 200 mL up to 1 or 2 L. Probably some may say that the equipment implies a very high initial investment, but it is really worth it in the long run, as the membranes are totally reusable and have a long-term working period. Millipore sells a wide range of UF cassettes (from 5 kDa to 300 kDa). Besides this, you can use the system in a concentration or purification (dialysis) mode.

Hope this helps. Regards.

As everyone say, it is better that you use ultra filtration at first if your protein will be not adsorbed with ultra filtration devices.

Dialysis using dialysis bags do not purify your protein to any significant extent. They can remove mostly small molecular weight compounds such as salts and small molecular weight peptides depending upon the nominal molecular weight cut-off of the dialysis membranes you use.