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The protein I am purifying needed an elution buffer with 2% Triton X-100. I formulated the elution buffer not keeping the CMC in mind. My goal is to make my protein sample triton free to check its stability and quantify its concentration. Lately, I learnt that micelles can be broken down upon dilution. So, I've diluted my protein sample below triton's CMC and tried concentrating it using Sartorius Vivaspin 5K (PES resin). I loaded 500 ul of diluted sample and was left with 100 ul after concentration using vivaspin. But, this sample gave an abnormal reading of above 3 OD after blanking against the buffer it is in. I presume, triton's interference is causing these discrepant readings.

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    $\begingroup$ You'll probably have to do multiple buffer exchanges if using a centrifugal filter. Dialysis is another option. $\endgroup$ – canadianer Jan 22 '15 at 16:13
  • $\begingroup$ This is a dialysed protein sample against Tris-HCl. Multiple buffer exchanges meaning, re-adding fresh buffer to the remain on filter? $\endgroup$ – piscean_1993 Jan 22 '15 at 16:27
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    $\begingroup$ If dialysis didn't remove it, I doubt filtration would. You can get beads specifically designed to remove Triton X-100 by adsorption. $\endgroup$ – canadianer Jan 22 '15 at 16:41

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