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Bio-UltraBio™ 一步法萤火虫萤光素酶检测试剂盒（即用型）
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为什么选择此级别
BioReagent,用于化学发光 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
避光,-80℃储存。特低温运输 。请查阅批次 COA 获取详细规格。
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文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
报告基因已被细胞生物学广泛使用以研究基因表达及其它与基因表达相关的细胞学事件，如受体活性、细胞内信号传导、mRNA 加工、蛋白质折叠及蛋白-蛋白相互作用等。萤火虫萤光素酶是基因表达中最常用的报告基因之一，是一种极度灵敏、响应快速且易于使用的报告基因，所催化的化学发光反应是测量基因表达最敏感的分析工具之一。
萤火虫萤光素酶（Firefly luciferase）是一种分子量约为61 kDa的蛋白，在ATP、镁离子和氧气存在的条件下，能够催化萤光素（luciferin）氧化成oxyluciferin，在luciferin氧化的过程中会发出波长为560 nm左右的生物萤光，该萤光可通过化学发光仪进行测定。
Bio-UltraBio™ 一步法萤火虫萤光素酶检测试剂盒（即用型）是无需洗涤或收集细胞的通过化学发光法直接测定细胞内萤火虫萤光素酶活性的高灵敏度、高信号稳定性的一步法检测试剂盒。该试剂盒提供即用型萤火虫萤光素酶检测试剂，按照与培养液等体积的比例加入到细胞培养板内反应5分钟，即可进行化学发光检测。本试剂盒使用灵活便捷、检测灵敏度高并且发光信号比较稳定，信号半衰期约2小时。
注意事项：
1、 萤火虫萤光素酶检测试剂应尽量避免多次反复冻融。建议第一次使用时适当分装，然后避光保存于-80℃。
2、 酶促反应对温度较为敏感，加样检测前需将细胞培养板和检测试剂平衡至室温。
3、 检测仪器选择：能够检测化学发光的仪器都适用本试剂盒的检测，但是针对相同的样品，不同检测器本底信号值和测量值均可能不同；且对于同一样本检测，不同仪器的数值不可横向比较。
4、 为防止孔间干扰，推荐使用不透明白色或黑色细胞培养板。
使用说明：
1. 细胞裂解
1) 贴壁细胞：不用吸除细胞培养基，直接加入与培养基等体积的检测试剂。
悬浮细胞：同贴壁细胞。
2) 推荐使用量：
器皿类型 | 96孔板 | 48孔板 | 24孔板 | 12孔板 | 6孔板 |
培养基体积 (μl/孔) | 100 | 150 | 200 | 300 | 500 |
检测试剂体积 (μl/孔) | 100 | 150 | 200 | 300 | 500 |
3) 室温条件下，在微型震荡器上或静置裂解5min。
2. 加样
96孔板样品可直接进行后续检测，其他类型样品每孔吸取200μl混合液（培养基+检测试剂）到检测板。
3. 检测
启动化学发光检测仪或具备化学发光功能的多功能酶标仪，根据仪器说明书及实验需求进行参数设置，随后上机读取信号值。
Introduction
Genetic reporters are used commonly in cell biology to study gene expression and other cellular events coupled to gene expression, such as receptor activity, intracellular signal transduction, mRNA processing, protein folding and protein:protein interactions. Firefly luciferase is one of the most commonly used reporter genes in gene expression. It is an extremely sensitive, fast-responding and easy-to-use reporter gene. The chemiluminescence reaction it catalyzes is one of the most sensitive analytical tools for measuring gene expression.
Firefly luciferase is a protein with a molecular weight of approximately 61 kDa. In the presence of ATP, magnesium ions and oxygen, it can catalyze the oxidation of luciferin into oxyluciferin. During the oxidation process of luciferin, bioluminescence with a wavelength of approximately 560 nm is emitted, and this fluorescence can be measured by a chemiluminescence analyzer.
Bio-UltraBio™ One-step Firefly Luciferase Reporter Gene Assay Kit (Ready-to-use） is a one-step assay kit for directly measuring the intracellular firefly luciferase activity through chemiluminescence without the need for cell washing or collection. It features high sensitivity and high signal stability. The kit provides ready-to-use firefly luciferase assay reagents. Just add them to the cell culture plate in an equal volume to the culture medium and react for 5 minutes, then chemiluminescence detection can be carried out. This kit is flexible and convenient to use, with high detection sensitivity and relatively stable luminescence signals. The signal half-life is approximately 2 hour.
Precautions
1. The luciferase detection reagent for fireflies should be avoided from repeated freezing and thawing as much as possible. It is recommended to appropriately aliquot the reagent for the first use and then store it in a dark place at -80°C.
2. The enzymatic reaction is temperature-sensitive. Both the cell culture plate and the detection reagent must be equilibrated to room temperature prior to assay initiation.
3. Detector Selection: Any instrument capable of measuring chemiluminescence is suitable for use with this kit. However, for identical samples, the background signal and measured values may vary between different detectors. Furthermore, readings obtained from different instruments for the same sample are not directly comparable.
4. To prevent well-to-well crosstalk, the use of opaque white or black cell culture plates is recommended.
Instruction for use
1. Cell lysis
1) Adherent cells: Do not aspirate the cell culture medium; directly add the detection reagent of the same volume as the medium.
Suspended cells: The same as adherent cells.
2) Recommended usage volume:
Culture Vessel | 96-well plate | 48-well plate | 24-well plate | 12-well plate | 6-well plate |
Culture Medium Volume (μl/well) | 100 | 150 | 200 | 300 | 500 |
Detection Reagent Volume (μl/well) | 100 | 150 | 200 | 300 | 500 |
3) Lyse the cells for 5 minutes at room temperature, either on a microplate shaker or by standing.
2. Sample Addition
Samples in 96-well plates may proceed directly to the subsequent assay. For other sample types, pipette 200 μl of the mixed solution (culture medium + detection reagent) per well into the assay plate.
3. Detection
Initiate the chemiluminescence detector or a multifunctional microplate reader equipped with chemiluminescence functionality. Set parameters according to the instrument manual and experimental requirements, then proceed to measure the signal values.
规格
别名
One-step萤火虫萤光素酶报告基因检测试剂盒 | 萤火虫萤光素酶报告基因检测试剂盒 | 萤光素酶检测系统
规格或纯度
BioReagent, 用于化学发光
英文名称
Bio-UltraBio™ One-step Firefly Luciferase Reporter Gene Assay Kit (Ready-to-use)
应用
Chemiluminescence
样品类型
cell or cell lysate
检测方法
Chemiluminescence
特异性
Firefly luciferase
检测仪器
酶标仪,化学发光仪
检测波长
560nm
敏感性
light-sensitive；Temperature-sensitive；Freeze-thaw sensitive
储存与运输
储存条件
避光,-80℃储存
运输条件
特低温运输
稳定性与储存
Store at -80℃ (12 months). Store in the dark.
组分与储存
技术文档
高级数据
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0434701 |
分析证书 | O1375169 |
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