I would also like to know the settings for FPLC system such as column flow rate, temperature, buffer to use and would like to know the standards which I have to use to develop molecular weight vs retention time.
Thanks Omar for your information. However, the manual you provided only discusses about protein purification and don't mention about carbohydrate purification using the column. Can I use the same flow rate for purification of carbohydrate from bacterial exopolysaccharide? I would be very grateful if you could provide me the details.
What is the expected size of the polysaccharide? Yes you can use the same flow rate.
Dear Omar, thanks for swift reply. The expected size of polysaccharide is more than 15kD and I will be using phosphate buffer pH 7 as eluent and temperature setting is 22'C. Do you think it will work? Also, what standards shall I use for the curve?
You should use dextrans as standards. You can use phosphate buffer and temperature set to 22 C.
Dear Omar, could you please explain me in detail because I don't understand why I can't use protein standards for calibration curve. Since I am using it for size exclusion and isn't molecules of same size will be eluted at the same time be it protein or polysaccharide?
That is for the same size and shape. Globular proteins are somewhat spherical while your polymer could be linear (do not know if this is true). Exclusion by the pores will be very different if you have a linear entity.
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