基本描述

别名

海肾萤光素酶报告基因检测试剂盒 | 荧光素酶报告基因

英文别名

Renilla Luciferase Assay Kit | Renilla Luciferase Glow Assay Kit

规格或纯度

BioReagent, 用于化学发光

稳定性与储存

-20℃储存（12个月），避光。

英文名称

Renilla Luciferase Reporter Gene Assay Kit (Standard)

储存温度

-20°C储存,避免反复冻融

运输条件

超低温冰袋运输

产品介绍

本产品是一种高灵敏度、稳定、均质的海肾萤光素酶报告基因检测试剂盒。本产品无需洗涤或收集细胞即可通过化学发光法直接测定细胞内海肾萤光素酶（sea pansy (Renilla reniformis) luciferase，简称 Renilla luciferase）活性。在细胞培养板内加入与培养液等体积的检测试剂反应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、测定样品的线性范围宽。另外，本产品也可以用于裂解并收集保存的细胞样品的检测。

本产品既可单独使用，直接裂解细胞检测细胞中海肾萤光素酶活性；也可与萤火虫萤光素酶活性检测试剂盒共同使用进行一体化的双萤光素酶检测。本试剂盒为即配即用型试剂，集细胞裂解、海肾萤光素酶检测于一体，通过“加入-混合-检测”免去了先裂解后检测的实验步骤，而且稳定的“辉光”更适合高通量样品的检测。

反应原理如下：

组分表

R1375759	Component	100T	1000T	Storage
R1375759A	Renilla萤光素酶检测缓冲液	10 mL	100 mL	-20℃. Store in the dark.
R1375759B	Renilla 萤光素酶检测底物 (100×)	100 μL	1 mL	-20℃. Store in the dark.

产品优势

1、本产品操作便捷，读数稳定，检测速度快：本产品集细胞裂解、海肾萤光素酶检测于一体，只需要加样-读板，即可完成实验。产生的光信号半衰期近乎1h，发光信号稳定。

2、使用灵活便捷，适用范围广：本产品发光信号稳定，既适用于少量样品的检测，也适用于大量样品的高通量筛选。

3、本产品可与萤火虫萤光素酶活性检测试剂盒共同使用：本产品可以抑制99.9%以上的萤火虫萤光素酶催化的发光信号，不仅可以单独检测细胞中的海肾萤光素酶活性，也可作为内参，与萤火虫萤光素酶活性检测试剂盒共同使用进行一体化的双萤光素酶检测。

4、兼容性强：本产品兼容各种常见培养液，正常培养液中的酚红、10%以内的胎牛血清或小牛血清、2%以内的DMSO或乙醇对信号和稳定性基本没有影响，常用的盐类或金属离子在正常浓度下也基本没有影响。
海肾萤光素酶报告基因产品对比

阿拉丁的三款海肾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非常高，可以立即进行检测，推荐使用Renilla萤光素酶报告基因检测试剂盒（闪光型）（货号S1375702）；

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Renilla萤光素酶报告基因检测试剂盒（标准型）（货号R1375759）；

如果对发光信号有一定要求，推荐使用Renilla萤光素酶报告基因检测试剂盒（辉光高亮型）（货号R1375756）。

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	是否可以搭配Firefly作为内参使用	是否为一步法可直接用于细胞检测
R1375759	Renilla萤光素酶报告基因检测试剂盒（标准型）	辉光型	高	灵敏	1h	一瓶溶剂一管底物	非常适合	是	是
S1375702	Renilla萤光素酶报告基因检测试剂盒（闪光型）	闪光型	非常高	非常灵敏	3-5min	一瓶裂解液一瓶溶剂一管底物	不适合	是	否
R1375756	Renilla萤光素酶报告基因检测试剂盒（辉光高亮型）	辉光型	很高	很灵敏	15-20min	一瓶裂解液一瓶溶剂一管底物	非常适合	否	是

使用说明：
一、自备材料

PBS；多道排枪；白色或黑色不透光细胞培养板；化学发光仪或酶标仪。

二、检测前准备

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、检测反应液配制：根据实际使用量，以100:1的比例将适量的Renilla萤光素酶检测缓冲液与Renilla萤光素酶检测底物（100×）混匀，室温避光备用。例如：如果需要使用100mL检测缓冲液，则需要加入1mL的检测底物。

注：建议现配现用，剩余的检测反应液当次实验结束后，直接舍弃，不要留存。

三、操作方法

1、细胞原位检测

(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：建议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) Renilla Luciferase反应检测

① 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海肾萤光素酶检测反应液，加样体积与初始细胞培养液体积相同并充分混匀。

Cell Culture Plate	6-well	12-well	24-well	48-well	96-well	384-well
含细胞的培养液	2mL	1mL	500μL	200μL	100μL	25μL
检测反应液	2mL	1mL	500μL	200μL	100μL	25μL

② 孵育5min后于化学发光仪或多功能酶标仪中检测RenillaLuciferase报告基因活性。
注：为得到最佳检测结果，请在加入检测反应液后1小时内完成检测。

2、裂解并收集保存的细胞样品检测

(1) 裂解细胞
① 贴壁细胞：吸去细胞培养基，用PBS缓慢轻柔地洗涤一次，洗去残留的培养基，然后将液体全部吸弃。

悬浮细胞：500-1000g转速离心去上清后，用PBS清洗细胞一次，再次离心收集细胞沉淀。

② 按照下表推荐加入适量的1×细胞裂解工作液（萤光素酶报告基因细胞裂解液 (通用型)，货号F900604），室温下静置或振动摇晃裂解15min，吹打并吸取全部细胞裂解产物至1.5mL离心管中，12000g离心5-10min，取上清用于后续检测。

Cell Culture Plate	6-well	12-well	24-well	48-well	96-well
1×细胞裂解工作液	500μL	200μL	100μL	50μL	20μL

注：若萤光素酶的表达水平过低，可适当减少细胞裂解工作液用量以提高蛋白浓度，但需保证裂解工作液体积可以覆盖细胞孔板底面，否则会影响裂解效果。

(2) Renilla Luciferase 反应检测

① 小心吸取20μL细胞裂解上清依次加入至检测管或酶标板中，再加入100μL平衡至室温的含有底物的海肾萤光素酶检测工作液。

② 孵育5min后于化学发光仪或多功能酶标仪中检测Renilla Luciferase报告基因活性。

注：如果对于数据的稳定性的要求不太高，可以在混匀后立即进行化学发光检测。
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、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。

5、Renilla 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。

6、海肾萤光素酶催化的生物发光的最强波长为480nm，请根据仪器要求设置相应的参数。

Renilla Luciferase Reporter Gene Assay Kit (Glow) is a highly sensitive, stable, and homogeneous Renilla Luciferase Reporter Gene Assay Kit. It enables the direct determination of intracellular sea pansy (Renilla reniformis) luciferase (Renilla luciferase) activity via chemiluminescence without the need for cell washing or harvesting. Simply add an equal volume of detection reagent to the culture medium in the cell culture plate, react for 5 minutes, and proceed with chemiluminescence detection. This kit offers flexible and convenient use, high detection sensitivity, and a wide linear range for sample measurement. Additionally, this product can also be used to detect lysed and stored cell samples.

This kit can be used alone to directly lyse cells and detect Renilla luciferase activity within cells. It can also be used in conjunction with a Firefly Luciferase Activity Assay Kit for integrated dual-luciferase detection. The kit features ready-to-use reagents that integrate cell lysis and Renilla luciferase detection into a single step. The "add-mix-detect" workflow eliminates the separate steps of lysis followed by detection, and the stable "glow-type" luminescence is particularly suitable for high-throughput sample screening.

Reaction Principle:

R1375759	Component	100T	1000T	Storage
R1375759A	Renilla Luciferase Assay Buffer	10 mL	100 mL	-20℃. Store in the dark.
R1375759B	Renilla Luciferase Assay Substrate (100×)	100 μL	1 mL	-20℃. Store in the dark.

Product Advantages:

Convenient Operation, Stable Reading, Fast Detection: This kit integrates cell lysis and Renilla luciferase detection. Simply add the reagent and read the plate to complete the experiment. The generated light signal has a half-life of nearly 1 hour, ensuring stable luminescence.
Flexible, Convenient, and Wide Applicability: The stable luminescent signal makes this product suitable for both small-scale sample detection and large-scale, high-throughput screening.
Compatible with Firefly Luciferase Assay Kits: This product suppresses over 99.9% of the luminescent signal catalyzed by firefly luciferase. It can not only be used alone to detect Renilla luciferase activity in cells but also serves as an internal control for integrated dual-luciferase detection when used with a Firefly Luciferase Activity Assay Kit.
High Compatibility: This product is compatible with various common culture media. Phenol red in standard media, fetal bovine serum (FBS) or calf serum up to 10%, DMSO or ethanol up to 2%, as well as common salts or metal ions at normal concentrations, have minimal impact on signal and stability.

Comparison of Renilla Luciferase Reporter Gene Products

The main features and differences among three Renilla Luciferase Reporter Gene Assay Kits from Aladdin are as follows:

For very high luminescent signal requirements and immediate detection: Recommended: Renilla Luciferase Reporter Gene Assay Kit (Flash-Type) (Cat. No. S1375702)
For both luminescent signal and stability requirements, suitable for batch or continuous operation: Recommended: Renilla Luciferase Reporter Gene Assay Kit (Standard Type) (Cat. No. R1375759)
For standard luminescent signal requirements: Recommended: Renilla Luciferase Reporter Gene Assay Kit (Glow Bright-Type) (Cat. No. R1375756)

Product Cat. No.	Product Name	Luminescence Type	Signal Intensity	Sensitivity	Half-Life	Packaging	Suitable for Batch Samples	Compatible for Firefly Co-detection	One-Step Cell Assay
R1375759	Renilla Luciferase Reporter Gene Assay Kit (Standard)	Bright	High	Sensitive	1h	1 bottle Buffer 1 vial Substrate	Very Suitable	Yes	Yes
S1375702	Renilla Luciferase Reporter Gene Assay Kit (Flash)	Flash	Very High	Highly Sensitive	3-5min	1 bottle Lysis Buffer 1 bottle Buffer 1 vial Substrate	Not Suitable	Yes	No
R1375756	Renilla Luciferase Reporter Gene Assay Kit (Glow Bright)	Bright	Very High	Highly Sensitive	15-20min	1 bottle Lysis Buffer 1 bottle Buffer 1 vial Substrate	Very Suitable	No	Yes

Instructions for Use:

I. User-Supplied Materials

PBS
Multichannel pipette
White or black opaque cell culture microplate
Luminometer or microplate reader.

II. Preparation Before Assay

Renilla Luciferase Assay Substrate (100×): Briefly centrifuge at low speed before each opening.
Preparation of Detection Reaction Solution: Mix Renilla Luciferase Assay Buffer and Renilla Luciferase Assay Substrate (100×) at a 100:1 ratio according to the actual required volume. Mix thoroughly and protect from light at room temperature. (e.g., For 100 mL Buffer, add 1 mL Substrate).
- Note: Prepare fresh for each use. Discard any unused reaction solution after the experiment. Do not store.

III. Procedure

1. In-situ Cell Assay
(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) Renilla Luciferase Reaction Detection
① Using a multichannel pipette, add the equilibrated Renilla Luciferase Detection Reaction Solution containing substrate to each cell culture well. The volume added should be equal to the initial volume of the culture medium in the well. Mix thoroughly.

Cell Culture Plate	6-well	12-well	24-well	48-well	96-well	384-well
Mediun With cells	2mL	1mL	500μL	200μL	100μL	25μL
Detection Reaction Solution	2mL	1mL	500μL	200μL	100μL	25μL

②Incubate for 5 minutes at room temperature, then measure Renilla Luciferase reporter gene activity using a luminometer or microplate reader.

Note: For optimal results, complete detection within 1 hour after adding the detection reaction solution.

2. Detection of Lysed and Stored Cell Samples
(1) Cell Lysis
① Adherent Cells: Aspirate the culture medium. Wash cells once gently with PBS to remove residual medium, then aspirate all liquid.
Suspension Cells: Centrifuge at 500-1000 × g to pellet cells. Aspirate supernatant. Wash pellet once with PBS, centrifuge again, and aspirate supernatant.
② Add the recommended volume of 1× Cell Lysis Working Solution (Luciferase Reporter Cell Lysis Buffer (Universal), Cat. No. F900604) as per the table below. Lyse by incubating at room temperature with gentle shaking or rocking for 15 minutes. Pipette to resuspend and transfer the entire lysate to a 1.5 mL microcentrifuge tube. Centrifuge at 12,000 × g for 5-10 minutes. Collect the supernatant for subsequent detection.

Cell Culture Plate	6-well	12-well	24-well	48-well	96-well
1× Cell Lysis Working Solution	500μL	200μL	100μL	50μL	20μL

Note: If luciferase expression levels are too low, the volume of Cell Lysis Working Solution may be reduced appropriately to increase protein concentration. Ensure the lysis solution volume is sufficient to cover the bottom of the well, otherwise lysis efficiency may be compromised.
(2) Renilla Luciferase Reaction Detection
① Carefully transfer 20 μL of cell lysate supernatant into assay tubes or a microplate well. Add 100 μL of equilibrated Renilla Luciferase Detection Working Solution containing substrate.
② Incubate for 5 minutes at room temperature, then measure Renilla Luciferase reporter gene activity using a luminometer or microplate reader.
*Note: If data stability requirements are less stringent, chemiluminescence detection can be performed immediately after mixing.

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.
Multiple Plate Assay: If detecting multiple cell culture plates simultaneously, strive to maintain consistent incubation time after adding the detection solution to each plate before reading. This ensures optimal and comparable results across plates.
Substrate Handling: Renilla Luciferase Assay Substrate (100×) is formulated in anhydrous ethanol. Due to the volatility of ethanol, the volume may appear significantly less than the labeled specification upon first opening. In this case, replenish the volume to the labeled specification using anhydrous ethanol, mix thoroughly, and use as directed.
Emission Wavelength: The bioluminescence catalyzed by Renilla luciferase exhibits a peak emission wavelength of 480 nm. Please set the instrument parameters accordingly.

名称和识别符

分子类型	试剂盒

化学和物理性质

敏感性	Light sensitive

安全和危险性（GHS）

SDS英文版

技术规格说明书

技术规格说明书

r1375759 renilla萤光素酶报告基因检测试剂盒标准型中文说明书.pdf

质检证书（CoA,COO,BSE/TSE 和分析图谱)

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

输入批号以搜索分析图谱:

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

x

体积

x

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）

x

体积 1 (V1)

=

浓度 2 (C2)

x

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

货号 (SKU)	包装规格	是否现货	价格	数量
R1375759-100T	100T	期货
R1375759-1000T	1000T	期货

Renilla萤光素酶报告基因检测试剂盒（标准型）

库存信息

库存信息

基本描述

名称和识别符

化学和物理性质

安全和危险性（GHS）

技术规格说明书

质检证书（CoA,COO,BSE/TSE 和分析图谱)

可替换产品

溶液计算器

摩尔浓度计算器

稀释计算器

复溶计算器