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。萤火虫荧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左右的生物荧光，该萤光可通过化学发光仪进行测定。

Bio-UltraBio™高亮版萤火虫萤光素酶分步检测试剂盒（即用型）是一种高灵敏度辉光型定量检测试剂盒，特别适用于萤火虫荧光素酶低表达的样品检测。该试剂盒提供即用型萤火虫萤光素酶检测试剂，只需将100μl萤火虫萤光素酶检测试剂与20μl~100μl细胞裂解样品混合后即可立即进行化学发光的检测，并且发光信号比较稳定，信号半衰期约30分钟。

产品组分表

B1373068	组分	100T	10 × 100T	储存条件	用量 / 次
B1373068A	5×报告基因细胞裂解液	10 mL	10×10ml	-80℃	100μL
B1373068B	萤火虫荧光素酶检测液	10 mL	10×10ml	-80℃, 避光保存。	100μL

注意事项：

1、报告基因细胞裂解液可在4℃短期存放，-20℃保存一年有效。

2、萤火虫萤光素酶检测试剂在-20℃保存其检测效果会逐渐下降，建议在1个月内尽快使用；-80℃避光保存，至少一年有效。

3、为保证萤光素酶检测试剂的稳定性，尽量避免多次反复冻融。建议第一次解冻后适当分装，然后避光保存于-20℃或-80℃。

4、萤光素酶检测试剂解冻后避免长时间暴露于室温。

5、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。

6、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。

7、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：吸尽培养液。

悬浮细胞：离心收集细胞，弃上清。

2) 根据培养器皿类型，参考下表加入适量1×Lysis Buffer (用无菌水按 4:1 稀释 A 组分)：

器皿类型	96孔板	48孔板	24孔板	12孔板	6孔板
裂解液体积 (μl/孔)	100	150	200	300	500

*注：若萤光素酶表达水平较低，可酌情减少裂解液用量（例如，6孔板可低至100μl/孔）。

3) 室温条件下，在微型震荡器上或静置裂解10min。

4）96孔板样品可直接进行后续检测，也可以先冻存，待以后再测定。冻存样品需融解，并达到室温后再进行测定。

5）其他类型样品10,000-15,000×g离心3-5分钟，取上清用于测定。

2. 试剂准备

融解萤火虫萤光素酶检测试剂，并平衡至室温。

3. 仪器设置

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。

4. 加样

取20-100μl细胞裂解样品进行检测（建议用量100μl；若样本量有限可减少用量，但同批次实验应保持一致）。同时，设置等体积裂解液作为空白对照。

5. 检测

向各孔中加入100μl萤火虫萤光素酶检测工作液，立即上机读取信号值。

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. 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™ Bright two-step Firefly Luciferase Reporter Gene Assay Kit (Ready-to-use）is a highly sensitive glow assay kit, particularly suitable for the detection of samples with low expression of firefly luciferase. This kit provides ready-to-use firefly luciferase detection solution. The chemiluminescence detection can be carried out directly after mixing 100μl of firefly luciferase detection reagent with 20μl to 100μl of cell lysis sample. The luminescent signal produced by the Bio-UltraBio™ Bright two-step Firefly Luciferase detection Reagent has a half-life of more than 30 minutes.

B1373068	Components	100T	10 × 100T	Storage	Quantity Per Test
B1373068A	Luciferase Assay Lysis Buffer	10 mL	10×10ml	-80℃	100μL
B1373068B	Firefly luciferase detection solution	10 mL	10×10ml	-80℃, Store in the dark.	100μL

Precautions

1. The reporter gene cell lysate can be stored for a short period at 4℃ and remains stable for up to one year when kept at -20℃.

2. The detection performance of firefly luciferase assay reagents may gradually decline when stored at -20°C. It is recommended to use them within one month. When stored at -80°C and protected from light, the reagents remain effective for at least one year.

3. To ensure the stability of the luciferase assay reagents, avoid repeated freeze-thaw cycles. It is advisable to aliquot the reagents after the first thaw and store them protected from light at -20°C or -80°C.

4. Prolonged exposure to room temperature should be avoided after the luciferase assay reagent has been thawed.

5. Enzymatic reactions are sensitive to temperature. Before adding samples for detection, the cell culture plate and the detection solution should be balanced to room temperature.

6. All instruments capable of detecting chemiluminescence are suitable for the detection of this kit. However, for the same sample, the background signal values and measured values of different detectors may vary. The values of different instruments cannot be compared directly when detecting the same sample.

7. To prevent interference between Wells, it is recommended to use opaque white or black cell culture plates.

Instruction for use

1. Cell lysis

1) Adherent cells: Aspirate the culture medium completely.

Suspension cells: Centrifuge to collect the cells and discard the supernatant.

2) Add an appropriate volume of 1× Lysis Buffer (prepared by diluting Component A 4:1 with sterile water) according to the culture vessel, as referenced in the table below:

Culture Vessel	96-well plate	48-well plate	24-well plate	12-well plate	6-well plate
Lysis Buffer Volume (μl/well)	100	150	200	300	500

*Note: If luciferase expression level is low, consider using less lysis buffer (e.g., as low as 100μl/well for a 6-well plate).*

3) Lyse the cells for 10 minutes at room temperature, either on a microplate shaker or by standing.

4) For 96-well plates: Lysates can be assayed directly or frozen for later analysis. Thaw frozen samples completely and equilibrate to room temperature before assaying.

5) For other vessel types: Centrifuge lysates at 10,000-15,000 × g for 3-5 minutes. Use the supernatant for the assay.

2. Reagent preparation

Thaw the Firefly Luciferase Detection Reagent and equilibrate it to room temperature.

3. Instrument Settings

Turn on the luminometer or a multifunctional microplate reader with chemiluminescence detection capability. Set instrument parameters according to the manufacturer's instructions and experimental requirements.

4. Sample Loading

Load 20-100 μl of cell lysate (or supernatant for centrifuged samples) for measurement (100 μl is recommended if sample volume permits; use a consistent volume across all samples in the same experiment). Use an equal volume of lysis buffer as a blank control.

5. Measurement

Add 100 μl of the Firefly Luciferase Detection Reagent to each sample. Mix thoroughly and measure the luminescent signal immediately.

规格

别名

增强型萤火虫萤光素酶检测试剂盒 | 萤光素酶检测系统

英文别名

Enhanced Firefly Luciferase Reporter Gene Assay Kit | Luciferase Reporter Gene Assay Kit | Luciferase Assay System

规格或纯度

BioReagent, 即用型, 用于化学发光

英文名称

Bio-UltraBio™ Bright two-step Firefly Luciferase Reporter Gene Assay Kit（Ready-to-use）

应用

Chemiluminescence

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

各组分在相应的储存条件下保质期1年。

技术文档

📋 安全数据表 (SDS)

全面的危险、操作、储存及法规合规文件。

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📊 产品数据表

产品规格和应用的快速参考摘要。

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🔬 规格说明书

该级别的完整质量属性和验收标准。

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高级数据

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C of A & Other Certificates(BSE/TSE, COO):

输入批号以搜索分析图谱:

通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！

找到1个结果

批号(Lot Number)	证书类型	货号
ZJ25F1230233	分析证书	B1373068

技术文档和文章

b1373068_bio-ultrabio bright two-step firefly luciferase reporter gene assay kit（ready-to-use） product manual.pdf

b1373068_bio-ultrabio高亮版萤火虫萤光素酶分步检测试剂盒即用型产品说明书.pdf

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

浓度

体积

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

体积 1 (V1)

浓度 2 (C2)

体积 2 (V2)

复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

质量（小瓶）

所需的复溶浓度

需要帮助选择级别？

我们的级别选择指南涵盖了目录中所有的变体的纯度、稳定剂状态和应用适用性。

查看 BioReagent 级别指南 → 查看 Ready-to-use 级别指南 → 查看 for Chemiluminescence 级别指南 →

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Bio-UltraBio™高亮版萤火虫萤光素酶分步检测试剂盒（即用型）