Green Glutaminyl Cyclase Activity Assay *Fluorimetric*
CAS号:
Green Glutaminyl Cyclase Activity Assay *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
绿色谷氨酰胺环化酶活性检测试剂盒 荧光法、Green Glutaminyl Cyclase Activity Assay *Fluorimetric*详细如下: Green Glutaminyl Cyclase Activity Assay Kit 提供了一种方便的两步均相程序,用于使用绿色荧光底物测量各种来源的酶活性。在第一步中,底物与谷氨酰胺环化酶或含酶样品一起孵育并转化为焦谷氨酸形式。其次,添加谷氨酰胺环化酶显影剂以去除 pGlu 残基并产生绿色荧光团。它可以在 490 nm 激发和 520 nm 发射进行检测。产生的荧光与酶活性成正比。该试剂盒是谷氨酰胺酰环化酶激活剂和抑制剂高通量筛选 (HTS) 的理想选择。绿色荧光团的长波长受细胞成分和测试化合物的自发荧光的干扰较小。检测灵敏度为 1.5 ng/mL 的活性酶。 许多生物活性肽和蛋白质以焦谷氨酸 (pGlu) 作为其第一个氨基酸。这种修饰保护它们免受氨基肽酶的降解,并且在许多情况下决定了蛋白质或肽的功能。然而,焦谷氨酸修饰有时可能会导致负面的副作用。 Add test compounds and enzyme solution to the microplate wells. For one well of a 96-well plate, suggested volume of Glutaminyl Cyclase enzyme solution is 40 uL and 10 uL of test compound. Establish the following control wells at the same time, as deemed necessary: Positive control contains Glutaminyl Cyclase enzyme without test compound. Inhibitor control contains enzyme and 1-BenzylImidazole. Vehicle control contains enzyme and vehicle used in delivering test compound (e.g. DMSO, concentration not to exceed 1%). Test compound control contains assay buffer (Component E) and test compound. Some test compounds may themselves be fluorescent or interfere with the Ex/Em wavelengths and thereby give false results. Substrate control contains assay buffer (Component E). Using the assay buffer (Component E), bring the total volume of all controls to 50 uL. Add 50 uL of Glutaminyl Cyclase substrate solution into each well, except the test compound control wells. For best accuracy, it is advisable to have substrate solution equilibrated to the assay temperature. Mix reagents completely by shaking the plate gently for no more than 30 sec. Cover and incubate plate for 30 minutes at 37 C°. Add 50 uL of the prepared Glutaminyl Cyclase developer and mix thoroughly. Incubate the plate for additional 30 min at 37 C°. Measure fluorescence signal at Ex/Em=490 nm/520 nm. An example of the inhibition of Glutaminyl Cyclase by 1-BenzylImidazole is shown in Figure 2.