植物组织
伞形酮
试剂
化学
孵化
色谱法
基质(水族馆)
荧光
水解
组织培养
生物化学
生物
植物
体外
香豆素
量子力学
物理
物理化学
有机化学
生态学
出处
期刊:CSH Protocols
[Cold Spring Harbor Laboratory]
日期:2007-02-01
卷期号:2007 (2): pdb.prot4688-pdb.prot4688
被引量:12
摘要
INTRODUCTION The activity of β-glucuronidase (GUS) can be accurately determined in intact plant tissue using 4-methylumbelliferyl β-D-glucuronide (4-MUG) as a substrate. Upon hydrolysis by GUS, the fluorochrome 4-methyl umbelliferone (4-MU) is produced. This method is based on the permeability of both 4-MUG and 4-MU through plant tissue. It consists of incubation of the tissue with the reagent and quantification of the fluorescence emitted by 4-MU in the solution. GUS activity in each sample can be normalized per unit tissue weight, per unit protein (if it is determined in the tissue after the incubation with 4-MUG), or per sample. The latter is particularly useful if GUS is expressed only in a subset of cells within the tissue assayed. The advantage of this method is that since many samples can be assayed with relatively little effort, it is particularly appropriate for large-scale screens. The following protocol has been adapted for the use of 96-well microtiter plates (200-μl wells).
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