: This chapter describes a method for the purification of urinary trypsin inhibitor (UTI), a small chondroitin sulfate proteoglycan with Ser-proteinase inhibitory activity, excreted at high levels into urine following an inflammatory condition. The method consists of two fractionation steps: an anion-exchange chromatography and a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Coomassie Brilliant Blue G-250 gel staining. Several UTI bands are excised from gel, minced, destained, and dehydrated for extraction with SDS-containing buffer, at 60 °C for 24 h. This allows for obtaining a highly purified UTI sample useful for both structural and functional studies.
A Method for Urinary Trypsin Inhibitor (UTI) Purification Combining Anion-Exchange Chromatography Enrichment and Preparative SDS-PAGE / Nieddu, Gabriele; Formato, Marilena; Lepedda, Antonio Junior. - 2619:(2023), pp. 239-248. [10.1007/978-1-0716-2946-8_17]
A Method for Urinary Trypsin Inhibitor (UTI) Purification Combining Anion-Exchange Chromatography Enrichment and Preparative SDS-PAGE
Nieddu, Gabriele;Formato, Marilena;Lepedda, Antonio Junior
2023-01-01
Abstract
: This chapter describes a method for the purification of urinary trypsin inhibitor (UTI), a small chondroitin sulfate proteoglycan with Ser-proteinase inhibitory activity, excreted at high levels into urine following an inflammatory condition. The method consists of two fractionation steps: an anion-exchange chromatography and a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by Coomassie Brilliant Blue G-250 gel staining. Several UTI bands are excised from gel, minced, destained, and dehydrated for extraction with SDS-containing buffer, at 60 °C for 24 h. This allows for obtaining a highly purified UTI sample useful for both structural and functional studies.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
