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Coelenterazine cp *UltraPure grade*

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Physical properties
Molecular weight415.48
SolventEthanol
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12352200

OverviewpdfSDSpdfProtocol


CAS
123437-25-2
Molecular weight
415.48
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 cp *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 mM240.685 µL1.203 mL2.407 mL12.034 mL24.069 mL
5 mM48.137 µL240.685 µL481.371 µL2.407 mL4.814 mL
10 mM24.069 µL120.343 µL240.685 µL1.203 mL2.407 mL

Molarity calculator

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Citations


View all 7 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)
The CLAVATA signaling pathway mediating stem cell fate in shoot meristems requires Ca2+ as a secondary cytosolic messenger
Authors: Chou, Hsuan and Zhu, Yingfang and Ma, Yi and Berkowitz, Gerald A
Journal: The Plant Journal (2016): 494--506
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
Teaching an old hormone new tricks: cytosolic Ca2+ elevation involvement in plant brassinosteroid signal transduction cascades
Authors: Zhao, Yichen and Qi, Zhi and Berkowitz, Gerald A
Journal: Plant Physiology (2013): 555--565

References


View all 150 references: Citation Explorer
Luminescence of imidazo[1,2-a]pyrazin-3(7H)-one compounds
Authors: Teranishi K., undefined
Journal: Bioorg Chem. (2006)
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Authors: Pfleger KD, Dromey JR, Dalrymple MB, Lim EM, Thomas WG, Eidne KA.
Journal: Cell Signal (2006): 1664
Crystal structure of obelin after Ca2+-triggered bioluminescence suggests neutral coelenteramide as the primary excited state
Authors: Liu ZJ, Stepanyuk GA, Vysotski ES, Lee J, Markova SV, Malikova NP, Wang BC.
Journal: Proc Natl Acad Sci U S A (2006): 2570
In vivo testing of Renilla luciferase substrate analogs in an orthotopic murine model of human glioblastoma
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Journal: Mol Imaging (2006): 57
Expression, purification and characterization of calcium-triggered luciferin-binding protein of Renilla reniformis
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Journal: Protein Expr Purif. (2006)
Transient expression of apoaequorin in zebrafish embryos: extending the ability to image calcium transients during later stages of development
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Self-illuminating quantum dots light the way
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Journal: Nat Biotechnol (2006): 326
Blue fluorescent protein from the calcium-sensitive photoprotein aequorin: catalytic properties for the oxidation of coelenterazine as an oxygenase
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Journal: FEBS Lett (2006): 1977
Mitochondrial adaptations within chronically ischemic swine myocardium
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In vivo RNA interference-mediated ablation of MDR1 P-glycoprotein
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Journal: Clin Cancer Res (2005): 4487