I need a dialysed FBS but the commercial one takes longer time. I saw buffer exchange protocols using 0.15M NaCl in Thermo or Sigma but it is difficult to maintain the exact volume.
Is it good idea to use centrifuge method using Amicon or Centricon to remove the amino acids from FBS? I think we can control the volume of FBS too and seems faster.
I haven't tried to do this, but it is my opinion that it would be a very slow process to use centrifugal ultrafiltration with FBS because of the high protein concentration. As the concentration proceeded, the viscosity of the FBS would increase, slowing the rate of filtration. To process a large amount of FBS this way would take a long time. Dialysis would be easier. Afterwards, you could restore the sample to its original volume by adding saline or, if the volume needed to be decreased, by a relatively brief use of the centrifugal ultrafilters..
Another approach might be to use tangential flow filtration, if you can get the equipment. It is a method that is suitable for rapid dialysis and diafiltration of large volume samples
Dear Dr. Adam, thank you for the answer. I will be doing in small scale - 10-20 ml. I am planning to dialyse by buffer exchange and restoring the volume by centrifugal ultrafiltration.
Hello, My question is: how many time is necesary to remove amino acids? using the 0.15M of NaCl.
If you start the dialysis first thing in the morning with a large volume of dialysis bath relative to the volume of FBS (at least 100X), change the dialysis bath with fresh solution late in the day, and dialyze overnight, you should get rid of nearly all the free amino acids. If you need really thorough removal, change the bath solution again and continue the dialysis for another day (always at 4oC). Dialysis is a slow process. If you need a faster process and/or need to process a larger volume of FBS, you should use diafiltration, such as tangential flow filtration, which requires special equipment.
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