Coelenterazine
(Synonyms: 腔肠素) 目录号 : GC17680
Coelenterazine 是多种荧光素酶的底物，例如 Renilla 荧光素酶 (Rluc) 和 Gaussia 荧光素酶 (Gluc)，也是水母发光蛋白的辅助因子。
Cas No.:55779-48-1
Sample solution is provided at 25 µL, 10mM.
Coelenterazine is a substrate of various luciferases such as Renilla luciferase (Rluc) and Gaussia luciferase (Gluc), and is also a cofactor of aequorin. It is widely used and can be used for detection Calcium concentration in living cells, gene reporter assays, BRET (Bioluminescence Resonance Energy Transfer) studies, ELISA, HTS, and chemiluminescence detection of ROS levels in tissues or cells[1].
In vitro, coelenterazine enabled the dynamic monitoring of the RAW 264.7 cell oxidative burst [2].Coelenterazine chemiluminescence did not depend on the activity of cell-derived myeloperoxidase.It is a very sensitive and specific superoxide anion chemiluminescent probe[3]. Coelenterazine is a more reliable probe, although susceptible to storage-dependent high background chemiluminescence, but does not require prereductio[4].
In vivo, there were early, preclinical, changes in the localization and magnitude of coelenterazine chemiluminescent foci[2]. Commercially available coelenterazine（soluble in DMSO） is a sensitive substrate for in vivo bioluminescence imaging of coelenterazine luciferase[5].
References:
[1]Jiang T, Du L, Li M. Lighting up bioluminescence with coelenterazine: strategies and applications. Photochem Photobiol Sci. 2016 Apr;15(4):466-80.
[2] Bronsart L, Nguyen L, et al. Reactive Oxygen Species Imaging in a Mouse Model of Inflammatory Bowel Disease. Mol Imaging Biol. 2016 Aug;18(4):473-8.
[3]Teranishi K. Non-invasive and accurate readout of superoxide anion in biological systems by near-infrared light. Anal Chim Acta. 2021 Sep 22;1179:338827.
[4] Kervinen M, P?tsi J, Finel M, Hassinen IE. Lucigenin and coelenterazine as superoxide probes in mitochondrial and bacterial membranes. Anal Biochem. 2004 Jan 1;324(1):45-51.?
[5]Lucas M, Solano F. 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.
Coelenterazine 是多种荧光素酶的底物，例如 Renilla 荧光素酶 (Rluc) 和 Gaussia 荧光素酶 (Gluc)，也是水母发光蛋白的辅助因子。用途广泛，可用于检测活细胞中的钙离子浓度、基因报告基因检测、BRET（生物发光共振能量转移）研究、ELISA、HTS，以及组织或细胞中ROS水平的化学发光检测[1]< /sup>.
在体外，腔肠素能够动态监测RAW 264.7细胞的氧化爆发[2]。腔肠素化学发光不依赖于细胞来源的髓过氧化物酶的活性。它是一种非常灵敏和特异的超氧阴离子化学发光探针[3].腔肠素是一种更可靠的探针，虽然对存储依赖性高背景化学发光敏感，但不需要预还原[4]。
在体内，腔肠素化学发光灶的定位和大小在早期、临床前发生了变化[2]。市售腔肠素（溶于DMSO）是腔肠素荧光素酶体内生物发光成像的敏感底物[5]。
| Cell experiment [1]: | |
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Cell lines |
RAW 264.7 cells |
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Preparation Method |
5 × 104 cRAW 264.7 cells were added to wells of a 96-well black plate in triplicate, cell counts were performed, and native coelenterazine was added to the sample (final coelenterazine concentration 0-100 μmol/l). Plates were imaged using an IVIS low-light imaging system |
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Reaction Conditions |
final coelenterazine concentration 0-100 μmol/l，60min |
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Applications |
RAW 264.7 chemiluminescent signal intensity positively correlates with coelenterazine concentration in culture. The production of chemiluminescence by RAW 264.7 cells in the absence of stimulation is consistent with the detection of superoxide anion production during oxidative phosphorylation. |
| Animal experiment [2]: | |
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Animal models |
10-26 weeks thirty-six mice (16 females, 20 males) |
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Preparation Method |
450 nl of virus (three sites) were injected into the cerebral cortex of mice, followed by coelenterazine injections at 28-day intervals (minimum = 26 days, maximum time = 74 days). Coelenterazine (dissolved in DMSO) and diluted to a concentration of 2.43 mM in sterile water (1 mg/ml) for cortical injection through a 34-gauge needle for imaging and electrophysiological recordings. |
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Dosage form |
0.2µl, 0.4µl and 1µl，the injection rate was 1.25 μL/min |
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Applications |
Coelenterazine strongly boosted activity in LMO3-expressing mice, photon production was proportional to emission rate, and maximal doses of coelenterazine moderately increased neural activity in naive mice and GLuc cohorts, which itself can be produced at high concentrations Nonspecific excitatory effects. |
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References: [1]. Bronsart L, Nguyen L, Habtezion A, Contag C. Reactive Oxygen Species Imaging in a Mouse Model of Inflammatory Bowel Disease. Mol Imaging Biol. 2016 Aug;18(4):473-8. [2]. Gomez-Ramirez M, More AI,et al. The BioLuminescent-OptoGenetic in vivo response to coelenterazine is proportional, sensitive, and specific in neocortex. J Neurosci Res. 2020 Mar;98(3):471-480. |
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| Cas No. | 55779-48-1 | SDF | |
| 别名 | 腔肠素 | ||
| 化学名 | 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 | ||
| 分子式 | C26H21N3O3 | 分子量 | 423.46 |
| 溶解度 | Ethanol : 2 mg/mL (4.72 mM; Need ultrasonic; DMSO can inactivate Coelenterazine's activity) | 储存条件 | -20°C, protect from light, stored under nitrogen,unstable in solution, ready to use. |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 2.3615 mL | 11.8075 mL | 23.615 mL |
| 5 mM | 0.4723 mL | 2.3615 mL | 4.723 mL |
| 10 mM | 0.2361 mL | 1.1807 mL | 2.3615 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
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Related Biological Data
Superoxide production by HEK293 cells stably expressing NOX5β and COS-7 cells transiently transfected with NOX5β in response to addition of hemin. B) luminescence of coelenterazine
Superoxide levels were also measured using coelenterazine luminescence.Cells were treated as described, but instead of L-012, 50 μM coelenterazine(GlpBio) was added to each well 10 min prior to read.
Redox biology 36 (2020): 101656. PMID: 32738790 IF: 11.793 -
Related Biological Data
Four crucial factors were optimized for the assay of the LIPS method. S/N was a ratio of the fluorescent value from a positive sample divided by that from a negative control.
The magnetic particles with the complex were suspended in 180 μL hGluc buffer, which consisted of 20 mM Tris-HCl (pH 7.4) and 100 mM NaCl and then the substrate of 5 μL 200 μM coelenterazine(GlpBio) were added to obtain a value of bioluminescence on a microplate reader.
Int J Biol Macromol (2024): 130964. PMID: 38499123 IF: 8.1996 -
Related Biological Data
Superoxide production by HEK293 cells stably expressing NOX5β and COS-7 cells transiently transfected with NOX5β in response to the NO donor SNAP. B) Superoxide production as measured by luminescence of coelenterazine after 1 hr incubation with DMSO control or 50 μM SNAP.
Radicicol, hemin, NO donor, or Fe(PPIX)NO were added at various timepoints and either 400 μM L-012 or 50 μM coelenterazine(GlpBio) was added 10 min prior to read.
Free Radical Biology and Medicine (2021). PMID: 34139309 IF: 7.373 -
Related Biological Data
Optimization of the RBD-hGluc LIPS assay conditions. Here, S/N, which means signal of the luminescence of the positive samples over that of a negative sample.
After thoroughly washing, 5 μL 200 μM coelenterazine (Glpbio) were then added in the suspended magnetic beads for the measurement of relative light unit using a Synergy TM H1 multi-mode microplate reader.
Analytica Chimica Acta (2022): 340633. PMID: 36464447 IF: 6.9111