Purification procedure of tissue kallikrein Rat kallikrein was purified from urine to apparent homogeneity by ultrafiltration and ion-exchange chromatography on a DEAE-Sepharose
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Supplement 1: Methods Purification procedure of tissue kallikrein Rat kallikrein was purified from urine to apparent homogeneity by ultrafiltration and ion-exchange chromatography on a DEAE-Sepharose FF column (GE Healthcare, Buckinghamshire, UK), affinity chromatography on an aprotinin (Sigma, MO, USA) coupled HiTrap NHS-activated Sepharose HP column (GE Healthcare) and gel filtration twice on a TSK gel G3000SXXL column (Tosoh Corporation, Tokyo, Japan) according to the modified methods. 2 The enzyme preparation migrated as a single band with an apparent molecular mass of 33 kDa on SDS-PAGE under reducing conditions and eluted as a single peak with an apparent molecular mass of 53 KDa on gel filtration chromatography. The purified rat urine kallikrein had a specific activity of 47.619 μmol/min/mg protein towards the synthetic substrate Pro-Phe-Arg-MCA (Peptide Institute, Osaka, Japan).
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