How do you dissolve Coelenterazine?

How do you dissolve Coelenterazine?

Coelenterazine h is soluble in organic solvents such as ethanol and methanol, which should be purged with an inert gas. The solubility of coelenterazine h in these solvents is approximately 0.5 mg/ml. Do not dissolve in DMSO as it may oxidize. Coelenterazine h is sparingly soluble in aqueous solutions.

What is Coelenterazine used for?

Coelenterazine H is a substrate for Renilla and Gaussia luciferase, and can be used for in vivo imaging with cells transfected with our RediFect Green RenLuc lentiviral particles, or when either of these luciferases is used as a reporter.

Is Coelenterazine a protein?

Coelenterazine-v ligated to Ca2+-triggered coelenterazine-binding protein is a stable and efficient substrate of the red-shifted mutant of Renilla muelleri luciferase. Anal Bioanal Chem. 2010 Oct;398(4):1809-17. doi: 10.1007/s00216-010-4106-9.

What is NanoLuc?

NanoLuc™ (Nluc) is a 19.1 kDa, ATP- independent luciferase that utilizes a novel coelenterazine analog (furimazine) to produce high intensity, glow-type luminescence.

How does Gaussia luciferase work?

Gaussia luciferase can be expressed in mammalian cells using commercially available reporter plasmids. This luciferase, which does not require ATP, catalyzes the oxidation of the substrate coelenterazine in a reaction that emits light (at 470 nm), and has considerable advantages over other reporter genes.

What is coelenterazine H?

Coelenterazine h is a synthetic derivative of native coelenterazine that exhibits 16-fold higher luminescence intensity (emission maximum ~466 nm; half-total time of 0.6-1.2 sec) than native coelenterazine.

What is luciferin substrate?

D-Luciferin is a substrate for the enzyme firefly luciferase. Chemiluminescence at 560 nm is generated when luciferin is oxidized by an ATP-dependent process, catalyzed by luciferase. Product.

How long does luciferase last?

The luminescence signal decays over the course of about 10 minutes of reaction time, although signal half-life may vary depending on luciferase expression levels. The light production resulting from the luciferase reaction leads to formation of suicidal adenyl-oxyluciferin at the enzyme surface.

What happens when you mix luciferin and luciferase?

In a luminescent reaction, two types of chemicals, called luciferin and luciferase, combine together. The luciferase acts as an enzyme, allowing the luciferin to release energy as it is oxidized. The color of the light depends on the chemical structures of the chemicals.

What’s the difference between luciferin and luciferase?

One is a luciferin, or a light-producing substance. The other is a luciferase, or an enzyme that catalyzes the reaction. In some cases, the luciferin is a protein known as a photoprotein, and the light-making process requires a charged ion to activate the reaction.

What is Coelenterazine H?

How do you inject luciferin?

(Luciferin is typically administered either intraperitoneally or intravenously.) Example: Inject with 10 µL of Luciferin stock solution per gram of body weight (normally ~200 µL for a 20 g mouse for a standard 150 mg/kg injection). 5. Wait 10-20 minutes before imaging for maximum luciferase signal plateau.

Coelenterazine h is one of several synthetic coelenterazine analogs that confer different Ca 2+ affinities and spectral properties on the aequorin complex. Like native coelenterazine, these five novel derivatives can be used to reconstitute the aequorin complex both in vivo and in vitro.

How should coelenterazine be stored before reconstitution?

Prior to reconstitution, the coelenterazine should be stored at − 20 °C. In addition, coelenterazine is poorly soluble in water and so stock solutions are normally prepared in methanol. In this form, coelenterazine is stable for ~ 3 months at − 20 °C.

Which coelenterazine is used in HTS screening assay for GPCR?

Coelenterazines h and cp has been used in HTS screening assay for GPCRs. Table 1. Volume of Ethanol needed to reconstitute specific mass of Coelenterazine h *CAS#: 50909-86-9* to given concentration.

What is the difference between apoaequorin and coelenterazine?

Aequorins containing the cp, f or h form of coelenterazine exhibit 10-20 times stronger luminescence than that of apoaequorin reconstituted with native coelenterazine. Coelenterazines h and cp has been used in HTS screening assay for GPCRs.