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 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 235.045 µL | 1.175 mL | 2.35 mL | 11.752 mL | 23.505 mL |
5 mM | 47.009 µL | 235.045 µL | 470.09 µL | 2.35 mL | 4.701 mL |
10 mM | 23.505 µL | 117.523 µL | 235.045 µL | 1.175 mL | 2.35 mL |
Molarity calculator
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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)
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)
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
Authors: Kim, Donghun and Simo, Ladislav and Park, Yoonseong
Journal: Journal of Experimental Biology (2014): 3656--3663
References
View all 150 references: Citation Explorer
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Authors: Cheung CY, Webb SE, Meng A, Miller AL.
Journal: Int J Dev Biol (2006): 561
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.
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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
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
Authors: Frangioni JV., undefined
Journal: Nat Biotechnol (2006): 326
Expression, purification and characterization of calcium-triggered luciferin-binding protein of Renilla reniformis
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Authors: Inouye S., undefined
Journal: Protein Expr Purif. (2006)
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