{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,19]],"date-time":"2025-10-19T15:28:56Z","timestamp":1760887736281,"version":"build-2065373602"},"reference-count":26,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2005,4,14]],"date-time":"2005-04-14T00:00:00Z","timestamp":1113436800000},"content-version":"vor","delay-in-days":2540,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Electrophoresis"],"published-print":{"date-parts":[[1998,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Proteolytic enzymes in human pure pancreatic juice (PPJ), which was collected by cannulating the main pancreatic duct using endoscopy, were investigated by two\u2010dimensional zymography (2\u2010DZ). 2\u2010DZ was carried out by combining isoelectric focusing (IEF) in the first dimension with sodium dodecyl sulfate\u2010polyacrylamide gel electrophoresis (SDS\u2010PAGE) in the second dimension, using gels containing casein or gelatin as a substrate for the proteolytic enzymes. After electrophoresis, the gels were incubated in Triton X\u2010100 followed by incubation at 37\u00b0C in Tris buffer (pH 8.5) containing CaCl<jats:sub>2<\/jats:sub>. By staining the gels with Coomassie Brilliant Blue (CBB R\u2010250), proteolytic enzymes were detected as clear spots and zones against a blue background. Proteinase inhibitors, such as a cysteine proteinase inhibitor (E\u201064), a metalloproteinase inhibitor (EDTA), and a serine proteinase inhibitor (Pefabloc\u00ae SC), were added to PPJ in order to determine the types of proteinases. In patients with pancreatic cancer, spots of molecular weight (<jats:italic>M<\/jats:italic><jats:sub>r<\/jats:sub>) 70 000 and isoelectric points (p<jats:italic>I<\/jats:italic>) 5.3\u20135.5 were clearly detected on the gels containing casein and gelatin, while these spots were not detected in the PPJ from healthy subjects. The proteolytic activities of these spots were strongly inhibited by EDTA and Pefabloc SC but not E\u201064. These results suggest that the spots of <jats:italic>M<\/jats:italic><jats:sub>r<\/jats:sub> 70 000 and p<jats:italic>I<\/jats:italic> 5.3\u20135.5 in PPJ of pancreatic cancer might be matrix metalloproteinase 2, which is a candidate for tumor\u2010associated proteinase. 2\u2010DZ proved to be a tool for analysis of proteolytic enzymes in PPJ and for the clinical diagnosis of pancreatic cancer.<\/jats:p>","DOI":"10.1002\/elps.1150190530","type":"journal-article","created":{"date-parts":[[2005,5,28]],"date-time":"2005-05-28T12:48:58Z","timestamp":1117284538000},"page":"782-787","source":"Crossref","is-referenced-by-count":11,"title":["Two\u2010dimensional zymography for analysis of proteolytic enzymes in human pure pancreatic juice"],"prefix":"10.1002","volume":"19","author":[{"given":"Seiji","family":"Kaino","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toshifumi","family":"Furui","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Satoko","family":"Hatano","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Miyuki","family":"Kaino","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kiwamu","family":"Okita","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kazuyuki","family":"Nakamura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,4,14]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1007\/BF02776015","volume":"13","author":"Harada H.","year":"1978","journal-title":"Gastroenterol. 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