7 ethoxyresorufin

Manufactured by AnaSpec

Sourced in United States

7-ethoxyresorufin is a fluorescent dye used as a substrate for the measurement of cytochrome P450 1A1 (CYP1A1) enzyme activity. It is a widely used reagent in biological research.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions) Cytochrome c, by Merck Group (2 mentions) Hek293t 17 cells, by American Type Culture Collection (2 mentions) Coelenterazine 400a, by GoldBio (2 mentions) Antibiotic antimycotic solution, by Thermo Fisher Scientific (2 mentions) Coelenterazine h, by Promega (2 mentions) Synergy 4 plate reader, by Agilent Technologies (1 mentions)

3 protocols using 7 ethoxyresorufin

Cytochrome P450 Activity Assay

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CYP1A1 and CYP2C19 activities were measured fluorescently using ethoxyresorufin-O-deethylase (EROD) and 3-cyano-7-ethoxycoumarin (CEC) assays, respectively. For the EROD assay, cells were exposed to 7-ethoxyresorufin (10 μM, AnaSpec, Fremont, CA, United States) and salicylamide (inhibitor of phase II metabolism of resorufin, 1.5 mM, VWR) for 4 h. For the CEC assay, cells were exposed to CEC (25 μM) for 4 h. CYP1A2, CYP2B6, CYP2C9, and CYP3A4 activities were measured using CYP P450-Glo^TM assays (Promega) according to the manufacturer’s instructions. Omeprazole (20 μM) was used to induce CYP1A2. Phenobarbital (1 mM) was used to induce CYP2B6. Rifampin (10 μM) was used to induce all other CYP enzymes. Fluorescent and luminescent reads were performed on a BioTek Synergy 4 plate reader.

Shoemaker J.T., Zhang W., Atlas S.I., Bryan R.A., Inman S.W, & Vukasinovic J. (2020). A 3D Cell Culture Organ-on-a-Chip Platform With a Breathable Hemoglobin Analogue Augments and Extends Primary Human Hepatocyte Functions in vitro. Frontiers in Molecular Biosciences, 7, 568777.

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Quantifying Protein-Protein Interactions

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Dulbecco’s Modified Eagle’s Medium (DMEM), phosphate buffered saline (PBS), fetal bovine serum (FBS), and Lipofectamine 2000 were purchased from Invitrogen (Eugene, OR). The plasmids used to generate BRET vectors (pGFP²-N1, pGFP²-N2, pRluc-N2, pRluc-N3) and the pGFP²-Rluc vector were obtained from BioSignal Packard (Waltham, WA). GFP² is a wild-type GFP that has been modified by a F64L substitution mutation which results in brighter fluorescence but similar excitation and emission spectra. Human embryonic kidney (HEK)-293T/17 cells were obtained from ATCC (Manassas, VA). Antibiotic-antimycotic solution was purchased from Life Technologies (Carlsbad, CA). Coelenterazine 400A was purchased from Gold Biotechnology (St Louis, MO), and coelenterazine h was purchased from Promega (Madison, WI). 7-Ethoxyresorufin was purchased from Anaspec (Fremont, CA), and cytochrome c was obtained from Sigma (St Louis, MO).

Connick J.P., Reed J.R., Cawley G.F, & Backes W.L. (2020). Heme oxygenase-1 affects cytochrome P450 function through the formation of heteromeric complexes: Interactions between CYP1A2 and heme oxygenase-1. The Journal of Biological Chemistry, 296, 100030.

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BRET Vectors in HEK-293T Cells

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Dulbecco's Modified Eagle's Medium (DMEM), phosphate buffered saline (PBS), fetal bovine serum (FBS), and Lipofectamine 2000 were purchased from Invitrogen (Eugene, OR). The plasmids used to generate BRET vectors (pGFP 2 -N1, pGFP 2 -N2, pRluc-N2, pRluc-N3) and the pGFP 2 -Rluc vector were obtained from BioSignal Packard (Waltham, WA). GFP 2 is a wild-type green fluorescent protein (GFP) that has been modified by a F64L substitution mutation which results in brighter fluorescence but similar excitation and emission spectra. Human embryonic kidney (HEK)-293T/17 cells were obtained from ATCC (Manassas, VA). Antibiotic-antimycotic solution was purchased from Life Technologies (Carlsbad, CA). Coelenterazine 400A was purchased from Gold Biotechnology (St. Louis, MO), and coelenterazine h was purchased from Promega (Madison, WI). 7-ethoxyresorufin was purchased from Anaspec (Fremont, CA), and cytochrome c was obtained from Sigma (St. Louis, MO).

Connick J.P., Reed J.R., Cawley G.F., & Backes W.L. (2020). Heme oxygenase-1 affects cytochrome P450 function through the formation of heteromeric complexes: Interactions between CYP1A2 and heme oxygenase-1.

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