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Protein reversed phase analysis.. linear or step gradients? - which is preferred and why?? (Jan/01/2008 )

Hi alll!
For protein reversed phase analysis.. linear or step gradients of the organic phase is better used?
Which of these is preferred and why??



I would start with linear gradient to see how complex is the sample and how separation should be optimized.

I may be wrong but I think linear gradient would give better separation of peaks...


The advantage of a step gradient is that it will give a more concentrated elution, potentially less pure.


you can step up to near where your protein of interest elutes (not too near or you will lose at least some of your protein) to remove contaminants in bulk then linear to separate your protein from others.