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Coelenterazine Catalog No.GC17680

luminescent enzyme substrate

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1mg
$65.00
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5mg
$232.00
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Sample solution is provided at 25 µL, 10mM.

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Protocol

Cell experiment:

Cells are plated at a density of 5×104 cells per well into 24-well plates and grown to 80-100% confluency. Just before imaging, media are changed to a colorless solution containing (in mM): 2.7 KCl, 139 NaCl, 8.1 Na2HPO4, 0.7 H2O, 1.5 KH2PO4, 1.8 CaCl2, 1 MgCl2 and 5.5 d-glucose. Cells are preincubated for 15 min in the absence or presence of Pgp modulator, after which Coelenterazine (final concentration of 470 nM) is added directly to the cells[1].

Animal experiment:

Mice are anesthetized with metofane or isoflurane before tail vein injection of Coelenterazine (4 μg/g) formulated from an ethanol stock diluted in sodium phosphate buffer (50 mM). Bioluminescence imaging is performed on the in vivo imaging system at 2, 6, 8, and 11 min after injection. Anesthesia is maintained during imaging by nose cone delivery of 2.5% isoflurane. After imaging, animals are killed by cervical dislocation; tumors are then harvested and weighed[1].

References:

[1]. Markova SV, et al, Coelenterazine-dependent luciferases. Biochemistry (Mosc). 2015 Jun;80(6):714-32.
[2]. Lucas M, et al. Coelenterazine is a superoxide anion-sensitive chemiluminescent probe: its usefulness in the assay of respiratory burst in neutrophils. Anal Biochem. 1992 Nov 1;206(2):273-7.
[3]. Pichler A, et al. In vivo RNA interference-mediated ablation of MDR1 P-glycoprotein. Clin Cancer Res. 2005 Jun 15;11(12):4487-94.
[4]. Volk-Draper LD, et al. Novel model for basaloid triple-negative breast cancer: behavior in vivo and response to therapy. Neoplasia. 2012 Oct;14(10):926-42.

Chemical Properties

Cas No. 55779-48-1 SDF Download SDF
Synonyms N/A
Chemical Name 8-benzyl-2-(4-hydroxybenzyl)-6-(4-hydroxyphenyl)imidazo[1,2-a]pyrazin-3(7H)-one
Canonical SMILES OC1=CC=C(CC(C2=O)=NC3=C(NC(C4=CC=C(O)C=C4)=CN32)CC5=CC=CC=C5)C=C1
Formula C26H21N3O3 M.Wt 423.46
Solubility ≥42.3mg/mL in DMSO Storage Store at -20°C
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request
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Background

Target: superoxide anion

IC50: 69 μM (rat hepatocytes cells)

The small molecule coelenterazine, a luminescent enzyme substrate, can react with the reactive oxygen species (ROS) superoxide anion and peroxynitrite and produce a chemiluminescent signal [1]. Therefore, coelenterazine is used for monitoring reporter genes in ELISA, BRET, and HTS techniques. Reactive oxygen species (ROS) play an important role in tumor development and survival and it is a new treatment strategy to target the cancer-specific ROS.

In vitro: Coelenterazine showed antioxidative properties in primary cultures of rat hepatocytes subjected to the oxidant tert-butyl hydroperoxide (t-BHP). Coelenterazine (3-10 μM) concentration-dependently increased survival and decreased lipid peroxidation in rat hepatocytes treated for 6 hr with t-BHP (250 μM) [2]. In addition, Coelenterazine (10 μmol/l) can be used to assay superoxide anion concentrations in cancer cell lines [1].

In vivo: the small molecule coelenterazine (5 mg/kg, intravenous administration) was a potential reporter of cancer-associated superoxide anion in the 4T1 breast adenocarcinoma mouse model [1]. Gli36-Rluc or Gli36-Gluc cells were injected into the leg muscle of nude mice. The mice were injected with 4 mg/kg of coelenterazine and imaged by a cooled charge-coupled device camera [3].

References:
1. Bronsart LL, Stokes C, Contag CH. Multimodality Imaging of Cancer Superoxide Anion Using the Small Molecule Coelenterazine. Mol Imaging Biol. 2016;18(2):166-71.
2. Dubuisson ML, de Wergifosse B, Trouet A, Baguet F, Marchand-Brynaert J, Rees JF. Antioxidative properties of natural coelenterazine and synthetic methyl coelenterazine in rat hepatocytes subjected to tert-butyl hydroperoxide-induced oxidative stress. Biochem Pharmacol. 2000;60(4):471-8.
3. Morse D, Tannous BA. A water-soluble coelenterazine for sensitive in vivo imaging of coelenterate luciferases. Mol Ther. 2012;20(4):692-3.