title: Silver-stained fibrin zymography: separation of proteases and activity detection using a single substrate-containing gel.
abstract: A new zymogram method, silver-stained fibrin zymography, for separation of protease bands and activity detection using a single substrate gel, was developed. The method takes advantage of the nanoscale sensitivity of both zymography and silver staining. After SDS-PAGE in a gel containing fibrin, the gel was incubated in enzyme reaction buffer and the zymogram was silver-stained. Bands with protease activity were stained with silver in clear areas where the protein substrate had been degraded. The molecular sizes of proteases were accurately determined. Furthermore, proteases of high molecular weight were clearly and sharply resolved.
authors.affiliation: Institute Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology, Jeonbuk, Korea.
authors.forename: Dong-Min
authors.initials: DM
authors.lastname: Chung
authors.forename: Ki Eun
authors.initials: KE
authors.lastname: Kim
authors.forename: Keug-Hyun
authors.initials: KH
authors.lastname: Ahn
authors.forename: Chan-Sun
authors.initials: CS
authors.lastname: Park
authors.forename: Dong-Ho
authors.initials: DH
authors.lastname: Kim
authors.forename: Hong Bum
authors.initials: HB
authors.lastname: Koh
authors.forename: Hyo Kon
authors.initials: HK
authors.lastname: Chun
authors.forename: Byung-Dae
authors.initials: BD
authors.lastname: Yoon
authors.forename: Hong Jib
authors.initials: HJ
authors.lastname: Kim
authors.forename: Min-Soo
authors.initials: MS
authors.lastname: Kim
authors.forename: Nack-Shick
authors.initials: NS
authors.lastname: Choi
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004792:Enzyme Precursors; D005337:Fibrin / Q000737:chemistry / Q000378:metabolism; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D016622:Silver Staining
chemical_list: D001426:Bacterial Proteins; D004792:Enzyme Precursors; D005337:Fibrin; D010447:Peptide Hydrolases