萤火虫萤光素酶报告基因检测试剂盒（辉光型）是一种辉光型定量检测试剂盒，具有高灵敏度和发光信号稳定的特点，可以满足高通量检测萤光素酶在哺乳动物细胞中的表达。本品发光信号强度大幅度提升，有效提高了检测灵敏度，可以用于检测ADCC Reporter Bioassay或细胞中萤火虫萤光素酶 (firefly luciferase) 报告基因的表达。
G1375755
| Component
| 10 mL | 100 mL
| Storage
| G1375755A
| Firefly Luciferase Reaction Buffer (Glow)
| 10 mL
| 100 mL
| -20℃. Store in the dark.
| G1375755B
| Firefly Luciferase Substrate (Glow)
| 1 vial
| 1 vial
| -20℃. Store in the dark. |
|
萤火虫萤光素酶报告基因产品对比
阿拉丁的三款一步法萤火虫萤光素酶报告基因检测试剂盒的主要特点和差异如下。
如果对发光信号要求很高，并在30分钟内即可进行检测，推荐使用萤火虫萤光素酶报告基因检测试剂盒（辉光高亮型）（货号B1375760）；
如果对发光信号和稳定性都有要求，需要批量或连续操作，推荐使用萤火虫萤光素酶报告基因检测试剂盒（货号F773892）；
如果对发光信号有一定要求，推荐使用萤火虫萤光素酶报告基因检测试剂盒（辉光型）（货号G1375755）。
对于常规的实验室检测，优先推荐使用萤火虫萤光素酶报告基因检测试剂盒（辉光高亮型）（货号B1375760）。
| 产品货号 | 产品名称 | 发光类型 | 信号强度 | 灵敏度 | 半衰期 | 包装 | 批量样品检测 | 试剂混合后-20℃存储最佳使用时间 | | F773892 | 萤火虫萤光素酶报告基因检测试剂盒
| 辉光型 | 较高 | 较灵敏 | 可达2h | 一瓶溶剂
一瓶底物粉末 | 非常适合 | 一个月 | G1375755
| 萤火虫萤光素酶报告基因检测试剂盒（辉光型）
| 辉光型 | 高 | 灵敏
| 25-30min | 一瓶溶剂
一瓶底物粉末
| 非常适合
| 一周 | B1375760
| 萤火虫萤光素酶报告基因检测试剂盒（辉光高亮型）
| 辉光型 | 很高 | 非常灵敏 | 25-30min
| 一瓶溶剂
一瓶底物粉末
| 非常适合
| 一周 |
|
使用说明：
1、自备材料
PBS；多道排枪；白色或黑色不透光细胞培养板；化学发光仪或具有发光检测模块的酶标仪。
2、检测前准备
(1) 首次使用时将 Firefly Luciferase Reaction Buffer (Glow) 一次性全部倒 Firefly Luciferase Substrate (Glow) 瓶中，充分混匀后，按使用需求进行分装，建议-70℃长期保存或者短期存放于-20℃不超过一个月，并尽快使用。
(2) 每次实验前将分装后冻存的检测试剂平衡至室温。
3、操作方法
(1) 从细胞培养箱中取出细胞培养板，放置5-15min，恢复至室温。
注：使用白色或黑色不透光的细胞培养板，减少孔间的信号干扰。
(2) 加入检测溶液
使用多道排枪向每个细胞孔中加入平衡至室温的含有底物的 Firefly Luciferase Reaction Buffer (Glow)，加样体积与细胞培养液体积相同。例如，96孔板通常加入80-100μL培养液，相应加入80uL-100μL检测溶液；384孔板通常加入20-30μL培养液，相应加入20-30μL检测溶液。
(3) 振荡混匀
为了使得细胞裂解充分，建议将细胞培养板放在振荡混匀仪或附带振板功能的仪器上，室温条件下，采用中高速振板5min。
注：建议振板混匀时间为5min，可根据细胞量进行适当调整，确保细胞充分裂解，得到稳定的发光检测结果。
(4) 检测
振板混匀后于化学发光仪或酶标仪中检测Firefly Luciferase报告基因活性。
注：为得到最佳检测结果，请在加入检测试剂后尽快完成检测。
注意事项：
1、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。为防止孔间干扰，推荐使用不透明白色或黑色细胞培养板。
2、由于发光信号会受到检测环境如培养基组分、温度等影响，所以应确保同组内不同样本检测条件一致。
3、酶促反应对温度较为敏感，加样检测前务必将检测试剂以及细胞培养板平衡至室温。
4、为保证萤光素酶检测试剂稳定性，适量分装后建议-70℃长期避光保存或者短期存放于-20℃不超过一个月，并尽快使用，尽量避免多次反复冻融。
5、如需同时检测多个细胞培养板，请尽量确保每个细胞板加入检测溶液后孵育时间一致，再进行数据读取，以此获得最佳的检测结果。
6、萤火虫萤光素酶催化的生物发光的最强波长为 560 nm。
物理外观：
G1375755A: Liquid
G1375755B: Solid
Firefly Luciferase Reporter Gene Assay Kit (Glow) is a glow-type quantitative detection kit characterized by high sensitivity and stable luminescent signals. It is suitable for high-throughput detection of firefly luciferase expression in mammalian cells. This product significantly enhances luminescent signal intensity, effectively improving detection sensitivity. It can be used to detect firefly luciferase reporter gene expression in ADCC Reporter Bioassays or cells.
G1375755
| Component
| 10 mL | 100 mL
| Storage
| G1375755A
| Firefly Luciferase Reaction Buffer (Glow)
| 10 mL
| 100 mL
| -20℃. Store in the dark.
| G1375755B
| Firefly Luciferase Substrate (Glow)
| 1 vial
| 1 vial
| -20℃. Store in the dark. |
|
he main features and differences among three One-Step Firefly Luciferase Reporter Gene Assay Kits from Aladdin are as follows:
For high luminescent signal requirements and detection within 30 minutes: Recommended: Firefly Luciferase Reporter Gene Assay Kit (Glow Bright) (Cat. No. B1375760)
For both luminescent signal and stability requirements, suitable for batch or continuous operation: Recommended: Firefly Luciferase Reporter Gene Assay Kit (Cat. No. F773892)
For standard luminescent signal requirements: Recommended: Firefly Luciferase Reporter Gene Assay Kit (Glow) (Cat. No. G1375755)
For routine laboratory detection, the Firefly Luciferase Reporter Gene Assay Kit (Glow Bright) (Cat. No. B1375760) is the recommended priority.
Product Cat. No.
| Product Name
| Luminescence Type
| Signal Intensity
| Sensitivity
| Half-Life
| Packaging
| Suitable for Batch Samples
| Optimal Use After Mixing & Storing at -20°C
| | F773892 | Firefly Luciferase Reporter Gene Assay Kit
| Glow
| Relatively High
| Relatively Sensitive
| Up to 2h
| 1 bottle Buffer
1 vial Substrate
| Very Suitable
| 1 Month
| G1375755
| Firefly Luciferase Reporter Gene Assay Kit (Glow)
| Glow
| High
| Sensitive
| 25-30min | 1 bottle Buffer
1 vial Substrate
| Very Suitable
| 1 Week
| B1375760
| Firefly Luciferase Reporter Gene Assay Kit (Glow Bright)
| Glow
| Very High
| Highly Sensitive
| 25-30min
| 1 bottle Buffer
1 vial Substrate
| Very Suitable
| 1 Week |
|
Instructions for Use:
1. User-Supplied Materials
2. Preparation Before Assay
(1) Initial Reconstitution: Upon first use, transfer the entire contents of the Firefly Luciferase Reaction Buffer (Glow) bottle into the Firefly Luciferase Substrate (Glow) vial. Mix thoroughly until the substrate is completely dissolved. Aliquot the reconstituted detection reagent as needed. For long-term storage, aliquot and store at -70°C. For short-term storage, store aliquots at -20°C for no longer than one month. Use reconstituted reagent promptly.
(2) Pre-assay: Before each experiment, equilibrate the aliquoted, frozen detection reagent to room temperature.
3. Procedure
(1) Equilibrate Plates: Remove the cell culture plate from the incubator and allow it to stand at room temperature for 5-15 minutes.
Note: Use white or black opaque cell culture plates to minimize signal interference between wells.
(2) Add Detection Reagent: Using a multichannel pipette, add the equilibrated Firefly Luciferase Reaction Buffer (Glow) containing the substrate to each cell culture well. The volume added should be equal to the volume of the culture medium in the well. For example:
* 96-well plate: Typically add 80-100 µL culture medium; add 80-100 µL detection reagent.
* 384-well plate: Typically add 20-30 µL culture medium; add 20-30 µL detection reagent.
(3) Mix to Lyse Cells: To ensure complete cell lysis, place the plate on an orbital shaker or an instrument with shaking capability. Shake at medium to high speed at room temperature for 5 minutes.
Note: A shaking time of 5 minutes is recommended. This can be adjusted based on cell density to ensure complete lysis and stable luminescent results.
(4) Detection: Following the shaking step, immediately measure firefly luciferase reporter gene activity using a luminometer or microplate reader.
Note: For optimal results, perform the detection as soon as possible after adding the detection reagent.
Precautions
Instrument Selection: Any instrument capable of detecting chemiluminescence is suitable for use with this kit. However, background signal levels and measured values for identical samples may vary between different detectors. Values obtained from different instruments for the same sample are not directly comparable. To prevent well-to-well interference, the use of opaque white or black cell culture plates is strongly recommended.
Consistent Conditions: Luminescent signals can be influenced by environmental factors such as medium components and temperature. Ensure consistent detection conditions across all samples within an experiment.
Temperature Sensitivity: The enzymatic reaction is temperature-sensitive. It is essential to equilibrate both the detection reagent and the cell culture plate to room temperature before adding the reagent and performing the assay.
Reagent Stability: To maintain the stability of the luciferase assay reagent, aliquot it appropriately after reconstitution. For long-term storage, store aliquots at -70°C protected from light. For short-term storage, store at -20°C for no longer than one month. Use aliquots promptly and avoid repeated freeze-thaw cycles.
Multiple Plate Assay: If detecting multiple cell culture plates simultaneously, strive to maintain consistent incubation time after adding the detection reagent to each plate before reading. This ensures optimal and comparable results across plates.
Emission Wavelength: The bioluminescence catalyzed by firefly luciferase exhibits a peak emission wavelength of 560 nm.
ZJ25F0926937
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