Renilla luciferase reporter gene detection kit (standard type)

Cat. No.: R1375759

GRADE & PURITY BioReagent, for Chemiluminescence

Descripción general

  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:

1. 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.

2. 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.

3. 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.

4. 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

1. Renilla Luciferase Assay Substrate (100×): Briefly centrifuge at low speed before each opening.

2. 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:

1. 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.

2. 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.

3. 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.

4. 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.

5. 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.

6. Emission Wavelength: The bioluminescence catalyzed by Renilla luciferase exhibits a peak emission wavelength of 480 nm. Please set the instrument parameters accordingly.

Almacenamiento y envío

Condiciones de almacenamiento de almacenamiento

Store at -20°C,Avoid repeated freezing and thawing

Enviado en

Ice chest + Ice pads

Estabilidad y almacenamiento

Store at -20°C long term (12 months). Store in the dark.