I am trying to have a better resolution during purification. Would running my sample using S-100 HR give a better result from an S-200 HR run?
Also, other than the fractionation range, what is the difference between the two columns?
S-100 is for purifying protein upto 100,000 and S 200 is for purifying 200,000. Above 100,000 I will use S-200 but if mw of my protein is 50,000 I will use S-100.
Hi Mahesh,
Would it mean that S100 has a better resolution than S200? Based on what I have found in the manual, the difference between the two was only the fractionation range.
Dear Ernest
theoretical resolution defined as theoretical number plates/cm is similar but the different separation range make the coloumn able to resolve well different protein range therefore if you need to separate a protein of 20kda from a protein of 50 kda the S-100 is certainly better than S-200 which is preferable for high mw proteins ( for example to separate 500 kda for 100kda ).
one way to improve a little your resolution is also reduce the injection volume of the sample.
If your protein is around 50kda you can also consider the supedex 75 26/60 which is more expensive but it has higher resolution respect an sepacryl and if is well maintained can work for years.
good luck
Mamuele
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