抗糖尿病活性及作用机构研究-嘉南药理大学图书馆.pdfVIP

理 : (I)- : 糖尿 類 □ ■ CNPH-94-01 行94 年1 1 94 年12 31 行 94 年 2 27 (I)- α-glucosidase Xanthine Oxidase 降糖尿 度 p-nitrophenyl glucopyranoside (pNPG)α-glucosidase p-Nitrophenol 度 Xanthine Xanthine Oxidase Uric Acid 度 年鶴 alpha-Glucosidase Xanthine Oxidase BC 精 50%度 I.C.50 52 µg/ml 行 ; α-glucosidase; Xanthine Oxidase; (I)- 數 alpha-glucosidase Xanthine Oxidase 降糖尿 鶴 alpha-glucosidase Xanthine Oxidase BC 精行 (fractionation) 料 I. : alpha-glucosidase Xanthine Oxidase Xanthine p-nitrophenyl glucopyranoside (pNPG) Sigma E. Merck II. : Lambda 25 UV/VIS Spectrometer (Perkin-Elmer) Microplate Reader (Model 680, Bio-Rad) III. : IV. 1. α-glucosidase α-Glucosidase (0.085 unit)(0.01–200 µg/mL)p-nitrophenyl glucopyranoside (pNPG)phosphate buffer 372mL 0.1M Na CO . 2 3 α-glucosidase415 nmp-nitrophenol度 行(不 )(不 ) 列 : %= [C-(A-B)]*100/C C: (不 ) A:度 B: (不 ) PBS 0.01 M (H 7.4) 100 µl α-glucosidase (0.0085 unit/µl) 10 µl PNPG (3 mg/ml) 100 µl Incubated at 37ºC for exact 30 min. Na CO (1 M) 40 µl to stop reaction 2 3 Measure Absorbance at 415 nm using Micro Plate Reader 2. Xanthine Oxidase A. PBS ( 0.2 M , PH 7.5 ) (1) 0.2 M NaH2PO4 NaH PO ‧H O 27.6 g NaH PO ‧2 H O 31.2 g 1000 ml 2 4 2 2 4 2 (2) 0.2 M Na2HPO4 Na HPO ‧H O 35.61 g Na HPO ‧12H O 71.6 g 1000 ml 2 4 2 2 4 2 (1) 870