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Coelenterazine f *UltraPure grade*

Calcium measurement is critical for numerous biological investigations. The aequorin complex comprises a 22 kD apoaequorin protein, molecular oxygen and the luminophore coelenterazine. When three Ca2+ ions bind to this complex, coelenterazine is oxidized to coelenteramide, with a concomitant release of carbon dioxide and blue light. The approximately third-power dependence of aequorin's bioluminescence on Ca2+ concentration allows the measurement of Ca2+ concentrations with a broad detection range from ~0.1 µM to >100 µM. Unlike fluorescent Ca2+ indicators, Ca2+-bound aequorin can be detected without illuminating the sample, thereby eliminating interference from autofluorescence. Native coelenterazine and its derivatives confer different Ca2+ affinities and spectral properties on the aequorin complex. Recombinant apoaequorin reconstituted with coelenterazine hcp is reported to have the best luminescence overall, with both a high quantum yield and a fast response time. However, intracellular reconstitution of aequorin from coelenterazine analogs can be relatively slow. 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.

Calculators

Common stock solution preparation

Table 1. Volume of Ethanol needed to reconstitute specific mass of Coelenterazine f *UltraPure grade* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM235.045 µL1.175 mL2.35 mL11.752 mL23.505 mL
5 mM47.009 µL235.045 µL470.09 µL2.35 mL4.701 mL
10 mM23.505 µL117.523 µL235.045 µL1.175 mL2.35 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

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Citations

View all 3 citations: Citation Explorer
Intracellular Ca2+ is important for flagellin-triggered defense in Arabidopsis and involves inositol polyphosphate signaling
Authors: Ma, Yi and Zhao, Yichen and Berkowitz, Gerald A
Journal: Journal of Experimental Botany (2017)
Bioluminescence Resonance Energy Transfer (BRET) Assay for Determination of Molecular Interactions in Living Cells
Authors: Harikumar, Kaleeckal G and Yan, Yan and Xu, Ting-Hai and Melcher, Karsten and Xu, H Eric and Miller, Laurence J
Journal: (2017)
Orchestration of salivary secretion mediated by two different dopamine receptors in the blacklegged tick Ixodes scapularis
Authors: Kim, Donghun and Simo, Ladislav and Park, Yoonseong
Journal: Journal of Experimental Biology (2014): 3656--3663

References

View all 150 references: Citation Explorer
Transient expression of apoaequorin in zebrafish embryos: extending the ability to image calcium transients during later stages of development
Authors: Cheung CY, Webb SE, Meng A, Miller AL.
Journal: Int J Dev Biol (2006): 561
Mitochondrial adaptations within chronically ischemic swine myocardium
Authors: McFalls EO, Sluiter W, Schoonderwoerd K, Manintveld OC, Lamers JM, Bezstarosti K, van Beusekom HM, Sikora J, Ward HB, Merkus D, Duncker DJ.
Journal: J Mol Cell Cardiol (2006): 980
Blue fluorescent protein from the calcium-sensitive photoprotein aequorin: catalytic properties for the oxidation of coelenterazine as an oxygenase
Authors: Inouye S, Sasaki S.
Journal: FEBS Lett (2006): 1977
Self-illuminating quantum dots light the way
Authors: Frangioni JV., undefined
Journal: Nat Biotechnol (2006): 326
Expression, purification and characterization of calcium-triggered luciferin-binding protein of Renilla reniformis
Authors: Inouye S., undefined
Journal: Protein Expr Purif. (2006)
Page updated on November 9, 2024

Ordering information

Price
Unit size
250 ug
1 mg
Catalog Number
2115221158
Quantity
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Physical properties

Molecular weight

425.45

Solvent

Ethanol

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC12352200

CAS

123437-16-1
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