title: RAMA casein zymography: Time-saving and highly sensitive casein zymography for MMP7 and trypsin.
abstract: To detect metalloproteinase-7 (MMP7), zymography is conducted using a casein substrate and conventional CBB stain. It has disadvantages because it is time consuming and has low sensitivity. Previously, a sensitive method to detect MMP7 up to 30 pg was reported, however it required special substrates and complicated handlings. RAMA casein zymography described herein is rapid, sensitive, and reproducible. By applying high-sensitivity staining with low substrate conditions, the staining process is completed within 1 h and sensitivity was increased 100-fold. The method can detect 10 pg MMP7 by using commercially available casein without complicated handlings. Moreover, it increases detection sensitivity for trypsin.
authors: Expert Laboratory for Life Environments (ELLE), Yokohama City University, Yokohama, Japan.
authors: Hidetaro
authors: H
authors: Yasumitsu
authors: Expert Laboratory for Life Environments (ELLE), Yokohama City University, Yokohama, Japan.
authors: Yasuhiro
authors: Y
authors: Ozeki
authors: Expert Laboratory for Life Environments (ELLE), Yokohama City University, Yokohama, Japan.
authors: Robert A
authors: RA
authors: Kanaly
mesh_terms: D002364:Caseins / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry*; D020783:Matrix Metalloproteinase 7 / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D012394:Rosaniline Dyes / Q000737:chemistry; D012680:Sensitivity and Specificity; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism*
chemical_list: D002364:Caseins; D007202:Indicators and Reagents; D012394:Rosaniline Dyes; C024757:Coomassie brilliant blue R; D014357:Trypsin; D020783:Matrix Metalloproteinase 7
keywords: Casein zymography; MMP7; RAMA stain; Rapid detection; Trypsin