Coelenterazine (Synonyms: Luciferin (Oplophorus), Preluciferin (Watasenia)) |
Catalog No.GC17680
One-Click Copy Product Info
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Coelenterazine is a natural, cell-permeable marine luciferin. Coelenterazine is a substrate for multiple luciferases or photoproteins. Coelenterazine is a probe for calcium ion and reactive oxygen species detection (Ex/Em=429/466nm).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 55779-48-1
Sample solution is provided at 25 µL, 10mM.
Coelenterazine is a natural, cell-permeable marine luciferin. Coelenterazine is a substrate for multiple luciferases or photoproteins. Coelenterazine is a probe for calcium ion and reactive oxygen species detection (Ex/Em=429/466nm). Coelenterazine can be used in reporter gene assays, in vivo bioluminescence imaging, Ca²⁺ detection, BRET and oxidative stress-related studies[1-4].
References:
[1] Markova SV, Vysotski ES. Coelenterazine-dependent luciferases. Biochemistry (Mosc). 2015 Jun;80(6):714-32.
[2] Jiang T, Song J, Zhang Y. Coelenterazine-Type Bioluminescence-Induced Optical Probes for Sensing and Controlling Biological Processes. Int J Mol Sci. 2023 Mar 7;24(6):5074.
[3] Krasitskaya VV, Bashmakova EE, Frank LA. Coelenterazine-Dependent Luciferases as a Powerful Analytical Tool for Research and Biomedical Applications. Int J Mol Sci. 2020 Oct 10;21(20):7465.
[4] Jiang T, Du L, Li M. Lighting up bioluminescence with coelenterazine: strategies and applications. Photochem Photobiol Sci. 2016 Apr;15(4):466-80.
Protocol for Coelenterazine use[1-3]
This protocol provides only a guide. Please modify according to specific needs.
Preparation of staining solution:
(1) Dissolve Coelenterazine in methanol to prepare a 4.7mM stock solution. Note: Aliquot unused stock solution and store at -20°C or -80°C protected from light. Avoid repeated freeze-thaw cycles.
(2) Dilute the stock solution with 0.1M potassium phosphate buffer to prepare a 0.4μM to 2.5μM working solution. Note: Adjust the working concentration according to actual conditions. Prepare fresh before each use.
Mitochondrial reactive oxygen species detection (using rat heart mitochondria as an example):
(1) Isolate rat heart mitochondria or submitochondrial particles (SMP). Adjust protein concentration to approximately 5mg/ml.
(2) Add 1 mg of mitochondria or SMP suspension to 2.5μM Coelenterazine and 2mg/ml NADH. Immediately measure cumulative luminescence signal within 15 seconds.
Cell staining:
(1) Culture cells expressing Renilla luciferase. Wash cells once with colorless solution.
(2) Add 0.4μM Coelenterazine. Incubate immediately at 37°C protected from light for 6 hours.
(3) Acquire images using a bioluminescence imaging system.
In vivo mouse imaging:
(1) Inject 2mg/kg Coelenterazine via tail vein into mice bearing xenograft tumors stably expressing Renilla luciferase.
(2) Allow Coelenterazine to circulate in vivo for approximately 5 minutes. Use a bioluminescence imaging system to capture images of orthotopic tumors and metastases. After sacrifice, homogenize major organs. Mix 50μL Coelenterazine substrate with 10μL tissue lysate. Quantify using a luminometer.
Precautions:
(1) Animal experiments must comply with animal welfare guidelines.
(2) For your safety and health, please wear a lab coat and disposable gloves during operation.
References:
[1] Volk-Draper LD, Rajput S, Hall KL, et al. Novel model for basaloid triple-negative breast cancer: behavior in vivo and response to therapy. Neoplasia. 2012 Oct;14(10):926-42.
[2] Raha S, Myint AT, Johnstone L, et al. Control of oxygen free radical formation from mitochondrial complex I: roles for protein kinase A and pyruvate dehydrogenase kinase. Free Radical Biology & Medicine. 2002;32(5):421-30.
[3] Pichler A, Zelcer N, Prior JL, et al. In vivo RNA interference-mediated ablation of MDR1 P-glycoprotein. Clin Cancer Res. 2005 Jun 15;11(12):4487-94.
| Cas No. | 55779-48-1 | SDF | |
| Synonyms | Luciferin (Oplophorus), Preluciferin (Watasenia) | ||
| 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 | Ethanol : 2 mg/mL (4.72 mM; Need ultrasonic; DMSO can inactivate Coelenterazine's activity) | Storage | -20°C, protect from light, stored under nitrogen,unstable in solution, ready to use. |
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.3615 mL | 11.8075 mL | 23.615 mL |
| 5 mM | 472.3 μL | 2.3615 mL | 4.723 mL |
| 10 mM | 236.1 μL | 1.1807 mL | 2.3615 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00% Appearance: A solid
- 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
Average Rating: 5 (Based on Reviews and 28 reference(s) in Google Scholar.)