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Bio-UltraBio™ 高亮版海肾萤光素酶一步检测试剂盒
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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
-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
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。海肾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。
海肾萤光素酶（Renilla luciferase）是一种分子量约为36 kDa的蛋白，在氧气存在的条件下，可以催化腔肠素（Coelenterazine）氧化成Coelenteramide。在Coelenterazine氧化的过程中，会发出波长为480nm左右的生物萤光，该萤光可通过化学发光仪进行测定。
Bio-UltraBio™ 高亮版海肾萤光素酶一步检测试剂盒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海肾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海肾萤光素酶检测试剂，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。本试剂盒发光强度比Bio-UltraBio™ 标准版海肾萤光素酶一步检测试剂盒提高了约2-6倍，最大可达 100万以上，信号半衰期约15-20分钟。
注意事项：
1、 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液平衡至室温。
2、 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。
3、 为防止孔间干扰，推荐使用不透明白色或黑色细胞培养板。
4、 海肾萤光素酶检测底物（100×）配制在无水乙醇中。由于无水乙醇容易挥发，有时会在初次使用时发现体积明显小于包装规格的情况，此时用无水乙醇把体积补足至包装规格，并混匀后即可使用。
5、 当酶表达量过低时，发光信号会降低，但信号稳定性会增强；当酶表达量过高时，信号半衰期会减少，但发光信号会增强。建议根据实际情况优化实验设计方案（如增加/减少质粒转染量），避免萤光素酶表达量过高/低。
使用说明：
1. 试剂准备
1） 海肾萤光素酶检测底物（100×）：每次开盖前需进行短暂低速离心
2） 检测试剂配制：根据实际使用量，以 99：1 的比例将适量的海肾萤光素酶检测缓冲液与海肾萤光素酶检测底物（100×）混匀，避光备用。
注：建议现配现用，剩余的检测试剂当次实验结束后，直接舍弃，不要留存。
2. 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：
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3)室温条件下，在微型震荡器上或静置裂解5min。
3. 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。
4. 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。
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. Renilla 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.
Renilla luciferase is a protein with a molecular weight of approximately 36 kDa. Under the presence of oxygen, renilla luciferase can catalyze the oxidation of Coelenterazine into Coelenteramide. During the oxidation process of Coelenterazine, bioluminescence with a wavelength of approximately 480 nm is emitted, and this fluorescence can be measured by a chemiluminescence analyzer.
Bio-UltraBio™ Bright one-step Renilla Luciferase Reporter Gene Assay Kit is a one-step assay kit for directly measuring the intracellular renilla 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 renilla 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. The luminescence intensity of this kit is approximately 2 to 6 times higher than that of the Bio-UltraBio™ Standard one-step Renilla Luciferase Reporter Gene Assay Kit. The maximum value can reach over 1 million. The signal has a half-life of approximately 15 to 20 minutes.
Precautions
1. 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.
2. 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.
3. To prevent well-to-well crosstalk, the use of opaque white or black cell culture plates is recommended.
4. Renilla luciferase substrate (100×) is prepared in anhydrous ethanol. Since anhydrous ethanol is prone to volatilization, sometimes it is observed that the volume is significantly smaller than the packaging specification during the initial use. In such cases, the volume can be replenished to the packaging specification using anhydrous ethanol, and then mixed well before use.
5. When enzyme expression is too low, the luminescent signal decreases but stability increases; when enzyme expression is too high, the signal half-life shortens but the luminescent signal intensifies. It is recommended to optimize the experimental design according to actual conditions (e.g., increasing or decreasing plasmid transfection amount) to avoid excessively high or low luciferase expression.
Instruction for use
1. Reagent preparation
1) Renilla luciferase substrate (100×): Before each opening of the container, a brief low-speed centrifugation is required.
2) Preparation of detection reagents: Based on the actual usage amount, mix an appropriate amount of Renilla luciferase detection buffer with the Renilla luciferase substrate (100×) in a 99:1 ratio. Store in the dark for later use.
Note: It is recommended to prepare the reagents immediately before use. The remaining detection reagents should be discarded at the end of the current experiment and not retained.
2. 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:
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3) Lyse the cells for 5 minutes at room temperature, either on a microplate shaker or by standing.
3. 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.
4.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
海肾萤光素酶报告基因检测试剂盒 | Renilla萤光素酶报告基因检测试剂盒（辉光高亮型） | 萤光素酶检测系统
英文别名
Renilla-Lumi™ Plus Luciferase Reporter Gene Assay Kit | Renilla Luciferase Reporter Gene Assay Kit (Glow Bright) | Luciferase Assay System
规格或纯度
BioReagent, 用于化学发光
英文名称
Bio-UltraBio™ Bright one-step Renilla Luciferase Reporter Gene Assay Kit
应用
Chemiluminescence
样品类型
cell or cell lysate
检测方法
Chemiluminescence
特异性
Renilla luciferase
检测仪器
酶标仪,化学发光仪
检测波长
480nm
敏感性
light-sensitive；Temperature-sensitive；Freeze-thaw sensitive
储存与运输
储存条件
-20°C储存
运输条件
超低温运输
稳定性与储存
各组分在相应的储存条件下保质期1年。
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|---|---|---|
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分析证书 | B1519760 |
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