Abstract
A short (75-mm) column of TSK-GEL G3000SW (S-column) can be used for both analytical and micropreparative gel chromatography. The elution conditions for the S-column were optimized and thereafter the column was well calibrated under native and denaturing conditions in a manner similar to that with a 600-mm column. The error in the Stokes radii of proteins obtained with the S-column was treated statistically. This S-column has the advantage of the economy in the time taken for analysis and solvent exchange and in the sample amounts required. Hence, it is useful for studying protein—ligand interactions by Hummel—Dreyer gel chromatography and for purifying S-carboxymethylated proteins prepared for microsequencing. These were exemplified by the analysis of the binding of o-nitrophenol to catechol 2,3-dioxygenase and of the NH 2-terminal amino acid sequence of rat pancreatic prophospholipase A 2.
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