一步法萤火虫萤光素酶报告基因检测试剂盒

One step firefly luciferase assay kit

O598309-500T

500T

避光,-80℃储存

超低温冰袋运输

产品介绍： 报告基因检测是现代分子生物学研究领域中分析结构基因旁侧区域潜在的顺式元件（如启动子、增强子和沉默子等）和反式作用因子相互作用关系的一种重要工具。 萤火虫萤光素酶被广泛应用于基因调控和药物筛选等方面。萤火虫萤光素酶是一种分子量约为61 kD的蛋白，在ATP、镁离子和氧气存在的条件下，可以催化萤光素生成氧化萤光素（Oxyluciferin），在萤光素被氧化的过程中，会产生光信号。 本试剂盒的光信号是一种瞬时光，在加入工作液后，需立即检测。光信号半衰期约为5 min。 使用方法： 1. 工作液配置 （1）将所有组分恢复至室温。 （2）用 A 组分充分稀释 B 组分(储液)，配制成 0.2 mg/mL 的萤火虫萤光素酶工作液，涡旋震荡，确保充分混匀。 注：萤火虫萤光素酶工作液不能反复冻融，若单次实验用量较少，建议按单次使用量分装。在室温下，工作液配置 3 h 后，活性下降约 10 %，5 h 后，活性下降约 25 %。 2. 化学发光值检测 （1）从培养箱中拿出细胞培养板，室温孵育 20 min，使其恢复至室温（22 – 25°C）。 （2）向培养板中加入与培养基等体积的萤火虫萤光素酶工作液，混匀。 （3）室温孵育 5 min。 注：孵育时间可随细胞类型和细胞数目适当调整。 （4）用多功能酶标仪或化学发光仪读取数值（仪器参数：测定时间为 10 s，测定间隔为 2 s）。 注意事项： 1. 使用前请将产品瞬时离心至管底，再进行后续实验。 2. 萤火虫萤光素酶催化的生物发光的最强波长为560 nm。 3. 为防止孔间干扰，建议使用白色不透光孔板。 建议： B 组分建议预先使用无菌水配置为 2 mg/mL 储液，A 组分及配置为储液的 B 组分，根据实验需求进行小批量分装。检测工作液建议现配现用，避免反复冻融。 组分： 一步法萤火虫荧光素酶检测缓冲液；D-荧光素； 应用范围： 主要用于ADCC检测 Product introduction： Reporter gene detection is an important tool for analyzing the interaction between potential cis elements (such as promoters, enhancers and silencers) and trans acting factors in the flanking region of structural genes in the field of modern molecular biology. Firefly luciferase is widely used in gene regulation and drug screening. Firefly luciferase is a protein with a molecular weight of about 61 KD. In the presence of ATP, magnesium ions and oxygen, it can catalyze the production of oxyluciferin from luciferin. In the process of luciferin oxidation, it will produce a light signal. The optical signal of this kit is a kind of instantaneous light, which needs to be detected immediately after adding the working solution. The half-life of optical signal is about 5 min. Instruction： 1.Working fluid configuration ( 1 ) Restore all components to room temperature. ( 2 ) The component B ( stock solution ) was fully diluted with component A to prepare a 0.2 mg / mL firefly luciferase working solution, which was vortexed and shaken to ensure full mixing. Note : The firefly luciferase working solution cannot be repeatedly frozen and thawed. If the dosage of a single experiment is small, it is recommended to subpackage according to a single dosage. At room temperature, the activity decreased by about 10 % after the working solution was configured for 3 h, and the activity decreased by about 25 % after 5 h. 2.chemiluminescence value detection ( 1 ) The cell culture plate was taken out from the incubator and incubated at room temperature for 20 min to restore it to room temperature ( 22-25 ° C ). ( 2 ) Add the same volume of firefly luciferase working solution with the medium to the culture plate and mix well. ( 3 ) Incubation at room temperature for 5 min. Note : The incubation time can be adjusted according to cell type and cell number. ( 4 ) The values were read by multifunctional microplate reader or chemiluminescence instrument ( instrument parameters : the determination time was 10 s, the determination interval was 2 s ). Matters needing attention： 1. please centrifuge the product to the bottom of the tube immediately before use, and then conduct subsequent experiments. 2. the strongest wavelength of bioluminescence catalyzed by firefly luciferase is 560 nm. 3. to prevent interference between holes, it is recommended to use white opaque orifice plate. Recommendation： Component B is recommended to use sterile water in advance to configure 2 mg / mL storage solution, A component and B component configured as storage solution, and small batch packaging according to the experimental requirements. The detection working fluid is recommended to be used now to avoid repeated freezing and thawing. Component： One-Step Firefly Luciferase Assay Buffer；D-Luciferin Scope of application： Mainly used for ADCC detection