520 颗粒酶 B 活性检测试剂盒,荧光法、520 Granzyme B Activity Assay Kit, Fluorimetric详细如下: 520 Granzyme B Assay Kit 为筛选酶抑制剂或使用新型内部猝灭的 5-FAM/QXL®FRET 底物连续测定 Granzyme B 活性提供了一种方便的检测方法。在被粒酶 B 切割后,FRET 底物将被分成两个片段,导致释放 5-FAM 荧光,可以在激发/发射 = 490 nm/520 nm 下对其进行监测。该试剂盒可用于检测生物样品和纯化酶制剂中颗粒酶B酶的活性。该检测可检测低至 0.1 ng/mL 的活性颗粒酶 B。 粒酶是结构相关的丝氨酸蛋白酶家族。它们储存在细胞毒性淋巴细胞的细胞毒性颗粒中。已知颗粒酶可诱导靶细胞凋亡。一旦进入靶细胞(病毒或肿瘤细胞),它们就有能力启动细胞死亡。颗粒酶 A 和 B 是研究最多的。 粒酶 B 是一种丝氨酸蛋白酶,主要由细胞毒性 T 淋巴细胞 (CTL) 和自然杀伤 (NK) 细胞分泌。它是半胱天冬酶依赖性细胞凋亡的主要介质之一。 Add test compounds and granzyme B enzyme solution (step 1.3) to the microplate wells. The suggested volume of enzyme solution for one well of a 96-well plate is 40 µL and test compound is 10 µL. Establish the following control wells at the same time, as deemed necessary Positive control: Add 40 µL granzyme B enzyme solution (step 1.3) and 10µL 1X assay buffer (step 1.1). Inhibitor control: Add 40 µL granzyme B enzyme solution (step 1.3) and 10µL granzyme B inhibitor (step 1.4). Vehicle control: Add 40 µL granzyme B enzyme solution (step 1.3) and 10µL vehicle used in delivering test compound (e.g. DMSO, concentration not to exceed 1%). Test compound control: Add 40µL 1X assay buffer (step 1.1) and 10µL test compound. Some test compounds have strong autofluorescence and may give false results. Substrate control: Add 50µL 1X assay buffer (step 1.1). The total volume of all controls should be 50 µL. Optional: Pre-incubate the plate for 10 min. at assay temperature. Any temperature (the assay temperature) from room temperature to 37°C may be used, as long as the subsequent incubations are performed at the same temperature.