Overview

Coelenterazine is a natural fluorescein that is abundant in nature and is the light-energy storage molecule of most Marine luminescent organisms (more than 75%). Coelenterazine can be used as a substrate for many luciferase enzymes, such as sea kidney luciferase (Rluc), Gaussia-secreted luciferase (Gluc), and Photoproteins including aequorin and Obelia. Unlike the firefly luciferin/luciferase system, the Coelenterazine/ luciferase system does not require adenosine triphosphate (ATP), making it easier to study biofluorescence in vivo. Therefore, Coelenterazine is commonly used as a luminescent substrate for fluorescence assays based reporter gene assays and live animal assays.

The main commercial Coelenterazine currently being used is Coelenterazine native. There are also many coelenterin derivatives such as Coelenterazine h, Coelenterazine 400a, Coelenterazine cp, Coelenterazine f, Coelenterazine hcp, Coelenterazine n equals are synthesized. Theoretically, these coelenterins can be used in the same experiments, but due to their differences in luminous wavelength, cell membrane permeability, and light quantum efficiency, they show different experimental effects in the same application.

Species of Coelenterazin

Coelenterazine Native

Also known as Coelenterazine free acid. It is the substrate of many luciferase enzymes such as Renilla luciferase (Rluc) and Gaussia luciferase (Gluc), and is also the cofactor of jellyfish luminescent protein.

Application scenarios: Detection of calcium ion concentration in living cells, gene report analysis, BRET (Bioluminescent resonance Energy transfer) study, ELISA, HTS and chemiluminescence detection of ROS levels in tissues or cells, etc.

Coelenterazine h

It is the substrate of many luciferase enzymes such as Renilla luciferase (Rluc) and Gaussia luciferase (Gluc), and is also the cofactor of jellyfish luminescent protein.

Application scenario: The luminous intensity is more than 10 times higher than Coelenterazine Native, which is suitable for reporter gene analysis and calcium ion concentration detection in living cells.

Coelenterazine hcp

It is one of the derivatives of Coelenterazine Native. The fluorescence intensity of Coelenterazine hcp-Aequorin complex is 190 times higher than that of Coelenterazine Native complex, with high quantum yield and fast Ca²⁺ reaction rate, which is very suitable for the detection of calcium ion levels.

Application scenario: Suitable for calcium ion level detection.

Coelenterazine 400a

Also known as DeepBlue CTM, it is one of the derivatives of Coelenterazine Native. It is a good substrate for Renilla luciferase (Rluc), and its emission wavelength is about 400nm, which has little interference with the signal of GFP receptor protein and cannot be oxidized by Gaussia secreted luciferase (Gluc).

Application scenario: It is the preferred Coelenterazine substrate for BRET (Bioluminescent resonance Energy Transfer) research.

Coelenterazine f

It is the preferred substrate for aequorin, and the only structural difference from Coelenterazine Native is that the hydroxyl group on its R-1 phenol group is replaced with fluorine (F). Compared to the total number of photons produced by the Coelenterazine Native-Apoaequorin complex, this complex produces only 80% of the photon energy.  The advantage is that it takes very little time to generate the aequorin complex.  When it comes into contact with Ca²⁺, it emits light quickly and in high yield, with a production intensity 20 times that of Coelenterazine Native.  In addition, it has good cell permeability.

Application scenario: When extremely high Ca²⁺ detection sensitivity is required to study jellyfish protein regeneration experiments, the use of substrates is recommended.

Coelenterazine cp

It is a derivative of Coelenterazine Native. The photoprotein complex formed with Apoaequorin protein is 15 times more luminous than Coelenterazine Native and has a faster Ca²⁺ reaction rate.

Application scenario: It can be used for high-throughput screening of G-protein-coupled receptors (GPCRs) drugs.

Coelenterazine n

Its fluorescence intensity is weaker in all Coelenterazine derivatives, and the reaction rate to Ca²⁺ is significantly lower than that of Coelenterazine Native.

Coelenterazine e

A derivative of Coelenterazine Native that is structurally one more ethyl group than Coelenterazine Native. As a substrate for Renilla luciferase, its fluorescence intensity is 137% that of Coelenterazine Native. Coelenterazine e has a high in vitro reactivity of jellyfish luminescent protein and dual emission peaks (405& 465), which enables the determination of Ca²⁺ concentration in the pCa5-7 range by the ratio of dual wavelengths. This method is independent of Coelenterazine concentration, thus improving the detection accuracy. Because the cell permeability of this derivative is poor, it does not have this advantage for intracellular related experiments.

Application scenario: Very useful substrate for Ca²⁺ low sensitivity detection.

Coelenterazine 2-methyl

It is a powerful cellular antioxidant. It can effectively act on intracellular reactive oxygen species, such as singlet oxygen and superoxide anions, and oxidative stress is an intermediate link in the process of apoptosis. In addition, the compound has the characteristics of non-toxicity and membrane permeability, so it can be used to study apoptosis.

Product Application:

1) As an important tool to study apoptosis;

2) It can also be used for chemiluminescence to detect superoxide anion and peroxynitrite anion levels.

Product information

Product name	Catalog number	Specifications
D-Luciferin,Sodium Salt	40901ES01/02/03/08	100mg/500mg/1g/5g
D-Luciferin,Potassium Salt	40902ES01/02/03/09	100mg/500mg/1g/5g
D-Luciferin Firefly,Free Acid	40903ES01/02/03	100mg/500mg/1g
Coelenterazine Native	40904ES02/03/08	1×500 μg/2×500 μg/5mg
Coelenterazine 400a	40905ES02/03	1×500 μg/2×500 μg
Coelenterazine h	40906ES02/03/08	1×500 μg/2×500 μg/5mg
Ready To Use Coelenterazine h	40907ES10	10 vials
Coelenterazine f	40908ES02/03	1×500 μg/2×500 μg

Published literature from users (incomplete statistics)

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