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Isoproterenol

Manufactured by Cayman Chemical
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Sourced in United States

Isoproterenol is a synthetic catecholamine, a beta-adrenergic agonist, with structural similarity to epinephrine. It is primarily used in laboratory research as a tool compound to study the effects of beta-adrenergic receptor activation.

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Lab products found in correlation

Glycerol standard solution, by Merck Group (2 mentions) Tgx protein gels, by Bio-Rad (2 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions) Polyethylenimine max pei, by Polysciences (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) 3h sb 269970, by PerkinElmer (1 mentions) N dodecyl β d maltoside ddm, by Anatrace (1 mentions) Cholesteryl hemisuccinate chs, by Anatrace (1 mentions) Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Digitonin, by Merck Group (1 mentions) Apyrase, by Merck Group (1 mentions) Gdp β s, by Merck Group (1 mentions) Protease and phosphatase inhibitor, by Cell Signaling Technology (1 mentions) Menthol, by Merck Group (1 mentions) Adenosine, by Merck Group (1 mentions)

6 protocols using isoproterenol

1

GPCR Ligand Binding Assay

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Trypsin, DPBS, PBS, FBS, MEM, DMEM, penicillin/streptomycin, and l-glutamine were obtained from Thermo Fisher Scientific. Receptor ligands (5-HT, isoproterenol, ICI-118,551, and MT) and forskolin were purchased from Cayman Chemical or MilliporeSigma. Detergents (n-dodecyl-β-d-maltoside [DDM] and cholesteryl hemisuccinate [CHS]) were obtained from Anatrace. Digitonin, apyrase, GDPβS, and GDP were purchased from MilliporeSigma or BioBasic. [3H]SB269970 was obtained from PerkinElmer, and polyethylenimine (PEI) MAX was purchased from Polysciences.
Jang W., Adams C.E., Liu H., Zhang C., Levy F.O., Andressen K.W, & Lambert N.A. (2020). An inactive receptor-G protein complex maintains the dynamic range of agonist-induced signaling. Proceedings of the National Academy of Sciences of the United States of America, 117(48), 30755-30762.
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2

Citrus Extract Naringenin Characterization

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Naringenin extract (NR) from whole citrus sinensis oranges (purity ≥ 30%) was purchased from GE Nutrients, Inc. (Gencor, Irvine, CA). BC, lycopene, and lutein were from Cayman Chemical Co. Protease and phosphatase inhibitors were purchased from Cell Signaling Technology (Danvers, MA), TGX protein gels from BIO-RAD (Hercules, CA). Type 1 collagenase, glycerol standard solution, adenosine, estradiol, human pituitary growth hormone, dobutamine hydrochloride, human atrial natriuretic peptide, ACTH, menthol, 8-CPT-cAMP were purchased from Sigma-Aldrich. Isoproterenol, human parathyroid hormone (1–34), CDCA, were purchased from Cayman Chemicals. Glycerol reagent A was from ZenBio (Durham, NC). All other chemicals were purchased from Sigma (St. Louis, Mo) unless otherwise indicated.
Primary antibodies used were UCP1 (#MAB6158, R&D Systems), GLUT4 (Ab654 Abcam), PGC-1α (ST1202, Sigma) and β-Actin (A5316, Sigma), or monoclonals from Santa Cruz against ATGL (sc-365278), adiponectin (sc-136131), PPARα (398394), PPARγ (sc-7273), NAMPT (sc-393444), RXRγ (sc-514134), RXRα (sc-515929). HRP-linked anti-rabbit (12-348, Sigma), anti-mouse (AP130P, Sigma) and anti-IgG kappa light chain (sc-516102, Santa Cruz) were used to detect specific antibody-antigen complexes. Western Lightning Plus-ECL was from PerkinElmer (Waltham, MA).
Coulter A.A., Greenway F.L., Zhang D., Ghosh S., Coulter C.R., James S.L., He Y., Cusimano L.A, & Rebello C.J. (2023). Naringenin and β-carotene convert human white adipocytes to a beige phenotype and elevate hormone- stimulated lipolysis. Frontiers in Endocrinology, 14, 1148954.
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3

Mitochondrial Respiration in Brown Adipocytes

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Primary brown adipocytes from different groups of mice were plated in seahorse plate and induced to mature brown adipocytes as described above. After differentiation, brown adipocytes were starved for 8 hrs, followed by treating the cells with or without 100 ng/mL IL18 for 24 h. Oxygen consumption rate (OCR) was performed on fully differentiated brown adipocytes (at day 8 of differentiation) with the Agilent Seahorse XF96 cellular Flux Analyzer (Agilent Technologies, Inc., Santa Clara, CA). Mitochondrial respiration was quantified using the Mito-stress test protocol. Four measurements were obtained under basal conditions and upon addition of isoproterenol (1 µM, 10009951, Cayman Chemical, Ann Arbor, MI), oligomycin (1 µg/mL, 75351), fluoro-carbonyl cyanide phenylhydrazone (FCCP, 1 µg/mL, C2920), rotenone (3 µM, R8875), and antimycin A (2 µg/mL, A8674) from Sigma-Aldrich. Non-mitochondrial respiration was subtracted to obtain basal, basal uncoupled, isoproterenol-stimulated uncoupled and maximal mitochondrial respiration.
Zhang X., Luo S., Wang M., Cao Q., Zhang Z., Huang Q., Li J., Deng Z., Liu T., Liu C.L., Meppen M., Vromman A., Flavell R.A., Hotamışlıgil G.S., Liu J., Libby P., Liu Z, & Shi G.P. (2022). Differential IL18 signaling via IL18 receptor and Na-Cl co-transporter discriminating thermogenesis and glucose metabolism regulation. Nature Communications, 13, 7582.
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4

Fucoxanthin Metabolism and Lipid Regulation

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Hyclone™ Dulbecco’s Modified Eagle Medium/Ham’s F12 (DMEM/F12 1:1) and Gibco™ fetal bovine serum (FBS) were purchased from ThermoFisher Scientific (Waltham, MA), Rosiglitazone was purchased from AK Scientific (Union City, CA). All other reagents used in the medium were purchased from Sigma-Aldrich (St. Louis, MO). Fucoxanthin (> 95% fucoxanthin, FN), bovine serum albumin (BSA), glycerol standard solution, and taurocholic acid were purchased from Sigma-Aldrich. Glycerol Reagent was purchased from Zenbio (The Triangle, NC). Isoproterenol was purchased from Cayman Chemicals (Ann Arbor, MI). An organic extract of the fucoxanthin plant containing 5wt% fucoxanthin (FEX) was provided by PLT Health Solutions, (Morristown, NJ). The purified alcohol of fucoxanthin, fucoxanthinol (FOL), was purchased from Wako Pure Chemical Industries (Osaka, Japan, 94% fucoxanthinol) and Sigma-Aldrich (≥ 97% fucoxanthinol). Cholesterol esterase was purchased from Worthington Biochemical Corporation (Lakewood, NJ). Protease and phosphatase were purchased from Cell Signaling Technology (Danvers, MA), RIPA buffer from Sigma, TGX protein gels from BIO-RAD (Hercules, CA).
Rebello C.J., Greenway F.L., Johnson W.D., Ribnicky D., Poulev A., Stadler K, & Coulter A.A. (2017). Fucoxanthin and its Metabolite Fucoxanthinol Do Not Induce Browning in Human Adipocytes. Journal of agricultural and food chemistry, 65(50), 10915-10924.
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5

Culturing HBEC6-KT and Calu-3 Airway Cells

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The human bronchial epithelial cell (HBEC6-KT) line was cultured in Keratinocyte Serum-Free Medium 1X (Gibco) according to the manufacturers directions and supplemented with P/S (VWR) at 1X70 (link). Human airway epithelial cell line Calu-3 (ATCC HTB-55) were cultured in Minimum Essential Medium Alpha Medium (Corning) and supplemented with Premium Grade FBS (VWR) at 10%, HEPES (Corning) at 1X, and P/S (VWR) at 1X. The following chemical reagents were dissolved in DMSO, cAMP elevating agents Forskolin (Cayman Chemical) and Isoproterenol (Cayman Chemical), ABCC4 inhibitors MK-571 (Cayman Chemical) and Ceefourin 1 (Abcam), PDE inhibitors Roflumilast (Cayman Chemical) and Rolipram (Cayman Chemical), and CFTR Inhibitor (Selleck Chemicals).
Nguyen J.P., Bianca M., Huff R.D., Tiessen N., Inman M.D, & Hirota J.A. (2021). Modulation of cAMP metabolism for CFTR potentiation in human airway epithelial cells. Scientific Reports, 11, 904.
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6

Adipocyte Differentiation Assay Protocol

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Dexamethasone, fetal bovine serum (FBS), high-glucose Dulbecco’s Modified Eagle’s Medium (DMEM), insulin solution, 3-isobutyl-1-methylxanthine, oil red O solution, and PhosSTOP were purchased from Sigma-Aldrich Corporation (St. Louis, MO, USA). Dimethyl sulfoxide (DMSO), ImmunoStar LD, lactate dehydrogenase (LDH)-Cytotoxic Test Wako, 10% neutral-buffered formalin, nobiletin, and sudachitin were purchased from FUJIFILM Wako Pure Chemical Corporation (Tokyo, Japan). The structures of sudachitin and nobiletin are shown in Figure 1A. Cyclic AMP ELISA Kit, H-89, isoproterenol, and SQ22536 were purchased from Cayman Chemical (Ann Arbor, MI, USA). cOmplete Protease Inhibitor Cocktail was purchased from Roche Diagnostics (Indianapolis, IN, USA). Block Ace was purchased from DS Pharma Biomedical Co., Ltd., (Osaka, Japan). EnzyChromTM Glycerol Assay Kit was purchased from BioAssay Systems (Hayward, CA, USA).
Abe T., Sato T, & Murotomi K. (2023). Sudachitin and Nobiletin Stimulate Lipolysis via Activation of the cAMP/PKA/HSL Pathway in 3T3-L1 Adipocytes. Foods, 12(10), 1947.
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