BioBM440 DEAE notes

BioBM440 DEAE notes - DEAE Ion exchange chromatography...

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DEAE Ion exchange chromatography involves the selective binding of charged proteins to a polymer matrix containing bound charged groups. In anion exchange chromatography, the matrix-bound groups are cation ic ( + charged) and thus selectively bind – charged proteins. Small cellulose particles cause low flow rates, because they get stuck in the interstices between the large particles. The more uniform the particle sizes , the higher the flow rate and the better the column functions. Allow particles to settle until next lab period. NOTE: This allows particles to settle into more homogeneous layers (larger at bottom, smaller at top), giving a higher flow rate. The final packed resin should fill ~½ column. Assume final column volume (packed resin) = 10mL . Arrange tubing from reservoir so that it dips below the top of the resin in the column and then back up to top. Thus when the reservoir runs dry, flow though the column stops. Whether
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