I was using Superose6 10/300 Gl column for purification of my target protein. the sample consists of various oligomeric state and i would like to identify the approximate elution volume where my target protein may elute during the size exclusion chromatography. I can't use any other techniques to identify it.
I am looking forward for the help!
Sebastian Schmitt
It is not ideal but Can i possibly use the elution profile of molecular weight standard present in the manual of the column to make a gesture?
Dear Meenal Chauhan
to provide you a better answer we need to see your elution profile. As Sebastian Schmitt
told you, the best approach is the SEC-MALS but in absence of it also the use of a MW standard could be a solution.The accuracy of this approach depends a lot from the MW of your protein and the resolution of your coloumn,
The idea to use the elution profile present in the manual can be valid if you have only 2 strong peaks with very different range, but is not accurate at all since the elution can change a little on the basis of the buffer, of the injection volume and on the protein properties (for example some proteins in a specific buffer can slighlty interact with the resin and the elution could be delayed respect than the expected ones)
best regards
Manuele
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