Coelentrazine

Manufactured by Promega

Sourced in United States

Coelentrazine is a substrate used in bioluminescence assays. It is a chemiluminescent compound that emits light when oxidized, typically in the presence of the enzyme luciferase. Coelentrazine is commonly used in cell-based reporter gene assays, bioimaging, and other applications that require sensitive detection of luminescent signals.

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Lab products found in correlation

Plasmid miniprep columns, by Qiagen (1 mentions) Horse anti mouse hrp linked antibody, by Cell Signaling Technology (1 mentions) Vent dna polymerase, by New England Biolabs (1 mentions) Gel extraction kit, by Qiagen (1 mentions) Ni nta agarose resin, by Qiagen (1 mentions) Alexa fluor 488 conjugated goat anti mouse antibody, by Thermo Fisher Scientific (1 mentions) Luminata forte western horseradish peroxidase hrp substrate, by Merck Group (1 mentions) Oligonucleotide, by Merck Group (1 mentions) Restriction enzyme, by New England Biolabs (1 mentions) Coelentrazine h, by Promega (1 mentions) Steadylite, by PerkinElmer (1 mentions) Prl cmv, by Promega (1 mentions) Cage 500, by Abberior (1 mentions) 552 n hydroxysuccinimide esters, by Abberior (1 mentions) Ha 11, by Fortrea (1 mentions) Ivis 200 imaging system, by PerkinElmer (1 mentions)

3 protocols using coelentrazine

Molecular Cloning and Protein Purification

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All chemicals used in the present study were of either analytical or molecular biology grades and were obtained from commercial sources. Media components were purchased from Difco (USA). Oligonucleotides were purchased from Sigma (India) and IDT (USA). Restriction enzymes, Vent DNA polymerase, and other DNA-modifying enzymes were obtained from New England Biolabs (USA). Gel-extraction kits, plasmid miniprep columns, and the Ni-NTA agarose resin were obtained from Qiagen (Germany). Hybridization nitrocellulose membrane (filter type 0.45 μm) and Luminata forte Western horseradish peroxidase (HRP) substrate was obtained from Millipore (India). Anti-His mouse monoclonal antibody (27E8) and horse anti-mouse HRP-linked antibody were procured from Cell Signaling Technology (USA). Anti-GSH mouse monoclonal antibody (ab19534) was from Abcam (UK). Alexa Fluor 488–conjugated goat anti-mouse antibody was obtained from Molecular Probes (USA). Coelentrazine was purchased from Promega (USA).

Chandel A., Das K.K, & Bachhawat A.K. (2016). Glutathione depletion activates the yeast vacuolar transient receptor potential channel, Yvc1p, by reversible glutathionylation of specific cysteines. Molecular Biology of the Cell, 27(24), 3913-3925.

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Recombinant Hormone Receptor Characterization

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Recombinant hCG and LH were purchased from National Peptides and Hormones Program (c/o A. F. Parlow, Harbor-UCLA Medical Center). For PALM studies, CAGE 500 and 552 N-hydroxysuccinimide esters for antibody conjugation and direct labeling of receptors were purchased from Abberior. Primary antibodies HA.11 and FLAG were purchased from Covance and Sigma, respectively. For BRET and pRL-cmv luciferase reporter assays, coelentrazine h and coelentrazine, respectively, were purchased from Promega. For cre-luc reporter gene assays, SteadyLite was purchased from PerkinElmer Life Sciences. Fluo-4 direct for calcium imaging was obtained from Invitrogen and HTRF-IP₁ assay from CisBio.

Jonas K.C., Fanelli F., Huhtaniemi I.T, & Hanyaloglu A.C. (2014). Single Molecule Analysis of Functionally Asymmetric G Protein-coupled Receptor (GPCR) Oligomers Reveals Diverse Spatial and Structural Assemblies. The Journal of Biological Chemistry, 290(7), 3875-3892.

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In vivo Luciferase Imaging of Particles

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Renilla luciferase production inside particles in vivo was monitored using IVIS 200 imaging system (PerkinElmer, Inc., Massachusetts, USA). Fifty µL of particles with luciferase-encoding DNA or without a DNA template were prepared and mixed with 10 µL of 170 µM of coelentrazine (Promega, Wisconsin, USA) - the enzyme’s substrate. The particles were kept on ice to eliminate in vitro protein production. The prepared mixtures of protein producing particles were injected intra-tumor to 10-week-old female BALB/c mice bearing a 4T1 cell-induced tumor. The experiment was carried 6 days after the tumor establishment. The mice were scanned in 10 minutes intervals for 1 hour, using the following IVIS scanning parameters: luminescence scan, working distance C, exposure time of 60 sec, large binning large and 1 f/s. During the experiment, the mice were anesthetized and their body temperature was kept using a red lamp.

Krinsky N., Kaduri M., Zinger A., Shainsky-Roitman J., Goldfeder M., Benhar I., Hershkovitz D, & Schroeder A. (2017). Synthetic Cell-like Particles Synthesize Therapeutic Proteins Inside Tumors. Advanced healthcare materials, 7(9), e1701163.

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