What is Coelenterazine H?
Coelenterazine h is one of several synthetic coelenterazine analogs that confer different Ca2+ 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 to dissolve coelenterazine?
Using coelenterazine / BRET 23 Coelenterazine 400a is hardly soluble in water, stock solution can be made in ethanol at 1mM (or ~0.4mg/ml) concentration and stored at –20°C in the dark. Avoid using DMSO to dissolve the material as it may cause oxidation.
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 frequently serves as a chromophore of photoproteins, such as aequorin, the first known photoprotein, isolated and characterized from the jellyfish Aequorea victoria (Shimomura et al., 1962).
Is Coelenterazine cell permeable?
Therefore it is not surprising that only one coelenterazine derivative (s-CTZ) with improved bioluminescence activity with GLuc has been reported. The compound structure has yet to be published, however, the published data strongly implies that s-CTZ is highly cell-permeable.
What is luciferin made of?
Bacterial luciferin is two-component system consisting of flavin mononucleotide and a fatty aldehyde found in bioluminescent bacteria.
Is luciferase an activity?
The luciferase assay is useful to study whether a protein of interest regulates a particular gene at the transcription level. When this protein activates transcription, the cell will produce luciferase enzyme. After the addition of a lysis buffer and a substrate, a luminometer quantifies the luciferase activity.
What is the drug luciferin?
Firefly luciferin (D-luciferin) is the luciferin, or light-emitting compound, found in many firefly (Lampyridae) species. The luciferin/luciferase system is used as a very sensitive reporter assay for gene expression. It can also be used for ATP assays in research applications or to detect bacterial contamination.
Why is it called luciferin?
“Luciferin” got its name from the Latin word “lucifer” (meaning “light-bearing”), which is also a source of the word that is sometimes used as a name of the devil.
Can luciferase be scanned?
The bioluminescence emitted can be detected and amplified using specialized cameras with highly sensitive detection systems , revealing the sites and levels of luciferase expression and activity from within a living animal.
What is the mechanism of action of coelenterazine?
Coelenterazine and its analogs are luminescent enzyme substrates for apoaequorin and Renilla luciferase. Apoaequorin is used as a calcium indicator whereas Renilla luciferase is commonly used as a reporter of transcription regulation. Coelenterazine binds to apoaequorin to form aequorin which emits light upon binding to calcium.
What is a 2-deoxy derivative of coelenterazine?
A 2-deoxy derivative of native Coelenterazine. Up to 20-fold higher initial light-output than native Coelenterazine with Apoaequorin. Reduced but longer lasting kinetic with Renilla luciferase. We recommend using CAT#399 NanoFuel Solvent for dissolving and stabilizing native Coelenterazine.
How do you prepare coelenterazine stock?
Coelenterazine stock (100 ×) is prepared in methanol and stored at − 80 °C protected from the light. It can be diluted to the working concentration of 5 μM by directly adding it to the wells where the cells are seeded.
What is the difference between coelenterazine and H-CTZ?
The photoprotein Aequorin is showing an up to 20-fold higher inital intensity with h-CTZ compared to native Coelenterazine. h-CTZ is also the preferred substrate for Renilla Luciferase in HTS (high throughput screens) due to its prolonged glow kinetic.