Cell culture medium and cell culture additives were from Life Technologies (Grand Island, NY). FuGENE HD transfection reagent and Promega GloSensor™ cAMP reagent were from Fisher Scientific (Pittsburgh, PA). FLIPR Calcium 5 Assay Kit was from Molecular Devices, Inc. (Sunnyvale, CA). Lipofectamine 2000 and TC-FlAsH™ II In-Cell Tetracysteine Tag Detection Kits were from Invitrogen (Grand Island, NY). hPTH(1–34) was obtained from Bachem, Inc. (Torrance, CA). Angiotensin II, [Arg8]-vasopressin, isoproternol, phenylephrine, and UK14303 were from Sigma-Aldrich (St. Louis, MO). Sphingosine 1-phosphate (S1P) was from Avanti Polar Lipids Inc. (Alabaster, AL). SI was from MP Biomedicals (Santa Ana, CA). SVdF and SBpA were synthesized by the Department of Pharmacology and Therapeutics, McGill University (Montreal, Quebec, Canada). Rabbit polyclonal anti-arrestin2/3 was a gift from Robert J. Lefkowitz (Duke University, Durham, NC). Anti-phospho-ERK1/2 IgG (T202/Y204; #9101) and anti-ERK1/2 IgG (#4695) were from Cell Signaling Technology (Beverly, MA). Horseradish peroxidase-conjugated donkey anti-rabbit IgG was from Jackson Immuno-Research Laboratories, Inc. (West Grove, PA).
Isoproternol
Manufactured by Merck Group
Isoproternol is a synthetic catecholamine that acts as a nonselective beta-adrenergic agonist. It is used in the laboratory setting for its pharmacological effects on the cardiovascular system.
Automatically generated - may contain errors
Lab products found in correlation
Angiotensin 2, by Merck Group (2 mentions)
Flipr calcium 5 assay kit, by Molecular Devices (2 mentions)
Cell culture medium, by Thermo Fisher Scientific (2 mentions)
Cell culture additives, by Thermo Fisher Scientific (2 mentions)
Tc flash 2 in cell tetracysteine tag detection kit, by Thermo Fisher Scientific (2 mentions)
Arg8 vasopressin, by Merck Group (2 mentions)
Uk14303, by Merck Group (2 mentions)
Anti phospho erk1 2 igg t202 y204 9101, by Cell Signaling Technology (2 mentions)
Anti erk1 2 igg, by Cell Signaling Technology (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Horseradish peroxidase conjugated donkey anti rabbit igg, by Jackson ImmunoResearch (2 mentions)
Phenylephrine, by Merck Group (2 mentions)
Sphingosine 1 phosphate (s1p), by Avanti Polar Lipids (2 mentions)
Fugene hd transfection reagent, by Promega (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Horse serum, by Merck Group (1 mentions)
Dmem high glucose, by Merck Group (1 mentions)
Collagen type 1, by Merck Group (1 mentions)
Penicillin streptomyocin, by Merck Group (1 mentions)
3 protocols using isoproternol
1
G Protein-Coupled Receptor Signaling Assays
2
Isolation and Culture of Murine Cardiomyocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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All animal work was approved by the University of Pittsburgh Division of Laboratory Animal Resources. Primary cardiomyocytes were isolated from Swiss Webster or Black 6 mouse neonates (P1–P3) as described (Ehler et al., 2013 (link); Wickline et al., 2016 (link)). For protein isolation, Swiss Webster-derived cardiomyocytes were plated onto 35 mm dishes (1×106 cells/dish) coated with Collagen Type I (Millipore). For immunostaining, cardiomyocytes were plated onto 35 mm MatTek dishes with 10 mm insets coated with Collage Type I. Cardiomyocytes were plated in plating media: 65% high-glucose DMEM (Thermo Fisher Scientific), 19% M-199 (Thermo Fisher Scientific), 10% horse serum (Thermo Fisher Scientific), 5% FBS (Atlanta Biologicals) and 1% penicillin-streptomycin (Thermo Fisher Scientific). Media was replaced 16 h after plating with maintenance media: 78% high-glucose DMEM, 17% M-199, 4% horse serum, 1% penicillin-streptomyocin, 1 µM AraC (Sigma) and 1 µM isoproternol (Sigma). Cells were cultured in maintenance media for 2–4 days until lysis or fixation.
3
G Protein-Coupled Receptor Signaling Assays
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