AFC 尿激酶 (uPA) 活性检测试剂盒 荧光法

库存 ：大量

英文名 ：AFC Urokinase (uPA) Activity Assay Kit *Fluorimetric*

CAS号 ：AFC Urokinase (uPA) Activity Assay Kit *Fluorimetric*

保质期 ：详见说明

供应商 ：齐源生物

保存条件 ：-20°C

规格 ：1 kit

AFC 尿激酶 (uPA) 活性检测试剂盒 荧光法、AFC Urokinase (uPA) Activity   Assay Kit *Fluorimetric*详细如下：
AFC 尿激酶 (uPA) 检测试剂盒针对酶抑制剂的筛选进行了优化。该试剂盒包含具有高反应性和低背景的荧光底物。uPA 切割底物，导致 AFC（7-氨基-4-三氟甲基香豆素）荧光团的释放。可以在激发/发射= 380/500 nm 处监测荧光。
尿激酶型纤溶酶原激活剂 (uPA) 是一种丝氨酸蛋白酶，其功能是将酶原纤溶酶原转化为具有活性的广谱丝氨酸蛋白酶纤溶酶。纤溶酶进而介导细胞外基质成分的细胞周围蛋白水解，并激活其他蛋白酶，例如导致细胞外基质进一步降解和重塑的基质金属蛋白酶和胶原酶。除了蛋白水解功能外，在与 uPA 受体 (uPAR) 结合后，uPA 还参与启动多种细胞内信号通路，这些信号通路通过与各种整合素和玻连蛋白的相互作用来调节细胞增殖、粘附和迁移。
1X Assay buffer: Add 5 ml of 2X assay buffer (Component C) to 5 mL deionized water.
uPA substrate solution: Dilute uPA substrate (Component A) 1:100 in 1X assay buffer according to Table 1. For each experiment, prepare fresh substrate solution.
uPA diluent: Dilute uPA enzyme (Component D) 1:100 in 1X assay buffer. This amount of enzyme is enough for a full 96-well plate. If not using the entire plate, adjust the amount of enzyme to be diluted accordingly.
uPA inhibitor (uPA-STOP^TM - synthetic uPA inhibitor): Dilute the 1 mM inhibitor solution (Component E) 1:10 to 100 uM in 1X assay buffer. Add 10 ul of the 100 uM inhibitor solution into each of the inhibitor control well of a 96-well plate.
Add test compounds and diluted enzyme solution to the microplate wells. For one well of a 96-well plate, the suggested volume of enzyme solution is 40 uL and 10 uL of test compound.
Simultaneously establish the following control wells, as deemed necessary:
Positive control contains the uPA without test compound.
Inhibitor control contains uPA and control inhibitor.
Vehicle control contains uPA and vehicle used in delivering test compound (e.g. DMSO, concentration not to exceed 1%).
Test compound control contains 1X assay buffer and test compound. Some test compounds have strong autofluorescence and may give false results.
Substrate control contains 1X assay buffer.
Using the 1X assay buffer, bring the total volume of all controls to 50 uL.
Optional: Pre-incubate the plate for 10 min at assay temperature. Any temperature (the assay temperature) from room temperature to 37 ℃ may be used, as long as the subsequent incubations are performed at the same temperature.