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- 详细信息
- 技术资料
- 库存：
大量
- 英文名：
520 Cathepsin L Assay Kit *Fluorimetric*
- CAS号：
520 Cathepsin L 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
520 组织蛋白酶 L 检测试剂盒 荧光法、520 Cathepsin L Assay Kit *Fluorimetric*详细如下：
520 组织蛋白酶 L 活性检测试剂盒使用 QXL™ 520/HiLyte™ Fluor 488 标记的 FRET 肽底物来测量酶活性。长波长 FRET 底物是根据 Cathepsin L 切割位点周围的序列设计的。QXL™ 520/HiLyte™ Fluor 488 对用于对完整底物进行最佳猝灭。当活性组织蛋白酶 L 切割 FRET 底物时，会导致 HiLyte Fluor™ 488 荧光增加，并在激发/发射 = 490 nm/520 nm 下进行监测。HiLyte™ Fluor 488 的荧光信号在低 pH 条件下是稳定的，这是组织蛋白酶 L 活性的最佳 pH 值。
组织蛋白酶是一类球状溶酶体蛋白酶，在哺乳动物细胞更新中起着至关重要的作用。它们降解多肽并通过它们的底物特异性来区分。组织蛋白酶 L 是一种溶酶体内肽酶，是半胱氨酸蛋白酶类木瓜蛋白酶家族的成员。
Set up the enzymatic reaction.
Add test compounds and diluted enzyme solution to the microplate wells. The suggested volume of enzyme solution for a 96-well plate is 40 uL/ well and test compound is 10 uL/well.
Simultaneously set up the following control wells, as deemed necessary:
Positive control contains the enzyme without test compound.
Inhibitor control contains Cathepsin L enzyme and inhibitor.
Vehicle control contains Cathepsin L enzyme and vehicle used in delivering test compound (e.g. DMSO, concentration not to exceed 1%).
Test compound control contains assay buffer and test compound. Some test compounds have strong autofluorescence and may give false results.
Substrate control contains assay buffer.
Using the assay buffer (with DTT) 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.
Run the enzymatic reaction.
Add 50 uL of Cathepsin L substrate solution into each well. For best accuracy, it is advisable to have the substrate solution equilibrated to the assay temperature. Mix the reagents completely by shaking the plate gently for 30 sec.
Measure fluorescence signal:
For kinetic reading: Immediately start measuring fluorescence intensity at Ex/Em=490 nm/520 nm continuously and record data every 5 min. for 30 to 60 min.
For end-point reading: Incubate the reaction for 30 to 60 min. Keep plate from direct light. Then measure fluorescence intensity at Ex/Em=490 nm/520 nm.
520 组织蛋白酶 L 活性检测试剂盒使用 QXL™ 520/HiLyte™ Fluor 488 标记的 FRET 肽底物来测量酶活性。长波长 FRET 底物是根据 Cathepsin L 切割位点周围的序列设计的。QXL™ 520/HiLyte™ Fluor 488 对用于对完整底物进行最佳猝灭。当活性组织蛋白酶 L 切割 FRET 底物时，会导致 HiLyte Fluor™ 488 荧光增加，并在激发/发射 = 490 nm/520 nm 下进行监测。HiLyte™ Fluor 488 的荧光信号在低 pH 条件下是稳定的，这是组织蛋白酶 L 活性的最佳 pH 值。
组织蛋白酶是一类球状溶酶体蛋白酶，在哺乳动物细胞更新中起着至关重要的作用。它们降解多肽并通过它们的底物特异性来区分。组织蛋白酶 L 是一种溶酶体内肽酶，是半胱氨酸蛋白酶类木瓜蛋白酶家族的成员。
Set up the enzymatic reaction.
Add test compounds and diluted enzyme solution to the microplate wells. The suggested volume of enzyme solution for a 96-well plate is 40 uL/ well and test compound is 10 uL/well.
Simultaneously set up the following control wells, as deemed necessary:
Positive control contains the enzyme without test compound.
Inhibitor control contains Cathepsin L enzyme and inhibitor.
Vehicle control contains Cathepsin L enzyme and vehicle used in delivering test compound (e.g. DMSO, concentration not to exceed 1%).
Test compound control contains assay buffer and test compound. Some test compounds have strong autofluorescence and may give false results.
Substrate control contains assay buffer.
Using the assay buffer (with DTT) 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.
Run the enzymatic reaction.
Add 50 uL of Cathepsin L substrate solution into each well. For best accuracy, it is advisable to have the substrate solution equilibrated to the assay temperature. Mix the reagents completely by shaking the plate gently for 30 sec.
Measure fluorescence signal:
For kinetic reading: Immediately start measuring fluorescence intensity at Ex/Em=490 nm/520 nm continuously and record data every 5 min. for 30 to 60 min.
For end-point reading: Incubate the reaction for 30 to 60 min. Keep plate from direct light. Then measure fluorescence intensity at Ex/Em=490 nm/520 nm.
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520 组织蛋白酶 L 检测试剂盒 荧光法
￥7056
