Isoproterenol, formoterol, GO6983, GO6976, RO32–0432, GF109203X, H89, PSB603, ESI-09, TPPB, NECA (adenosine-5′-N-ethyluronamide), MRS5698 and MRS2365 ([[(1R,2R,3S,4R,5S)-4-[6-amino-2-(methylthio)-9H-purin-9-yl]-2,3-dihydroxybicyclo[3.1.0]hex-1-yl]methyl] diphosphoric acid monoester trisodium salt) were purchased from Tocris (Ellisville, MO). CGS21680">CGS21680 , carbachol, cholera toxin (CTX) and PTX were from Sigma (St. Louis, MO). YM254890, enzastaurin, LY333531, VTX-27 and AB928 were purchased from MedChemExpress (Monmouth Junction, NJ, USA). BAY60-6583 (LUF6210, termed hereafter ‘BAY’) was synthesized at Leiden/Amsterdam Center for Drug Research and was provided by Ad IJzerman (Leiden, The Netherlands). All compounds were dissolved in DMSO except that CTX and PTX were in water, and proper controls were included in all experiments. AlphaScreen cAMP kit, SureFire p-ERK1/2 (Thr202/Tyr204) Assay Kit and AlphaScreen SureFire p-Akt 1/2/3 (p-Ser473) Assay Kit were purchased from PerkinElmer (Waltham, MA). Gs-null and Gq/11-null HEK293 cells were generated at Tohoku University, Sendai, Japan. HEK293 human embryonic kidney, PC-3 human prostate cancer, NIH-3T3 mouse fibroblast, and H9C2 rat cardiomyoblast cells were from ATCC (Manassas, VA); all other reagents were from standard commercial sources and of analytical grade.
Formoterol
Manufactured by Bio-Techne
Sourced in United Kingdom
Formoterol is a laboratory reagent used in cell culture and biochemical research applications. It functions as a beta-2 adrenergic receptor agonist, which can be utilized to study signaling pathways and cellular responses related to this receptor. The core function of Formoterol is to activate the beta-2 adrenergic receptor, enabling researchers to investigate its effects on cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Poly d lysine, by Merck Group (2 mentions)
Propranolol, by Merck Group (2 mentions)
Salmeterol, by Merck Group (2 mentions)
Ici 118 551, by Merck Group (2 mentions)
Nih 3t3 mouse fibroblast, by American Type Culture Collection (1 mentions)
Cgs 21680, by Merck Group (1 mentions)
Carbachol, by Merck Group (1 mentions)
Enzastaurin, by MedChemExpress (1 mentions)
Isoproterenol, by Bio-Techne (1 mentions)
Psb603, by Bio-Techne (1 mentions)
Alphascreen camp kit, by PerkinElmer (1 mentions)
Go6976, by Bio-Techne (1 mentions)
Ro 32 0432, by Bio-Techne (1 mentions)
Esi 09, by Bio-Techne (1 mentions)
Cholera toxin ctx, by Merck Group (1 mentions)
Ym254890, by MedChemExpress (1 mentions)
Ly333531, by MedChemExpress (1 mentions)
Ab928, by MedChemExpress (1 mentions)
Surefire p erk1 2 thr202 tyr204 assay kit, by PerkinElmer (1 mentions)
Alphascreen surefire p akt 1 2 3 p ser473 assay kit, by PerkinElmer (1 mentions)
7 protocols using formoterol
1
Characterization of Signaling Pathways
2
Formoterol Impacts Microglia Inflammation
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Primary microglia cells from C57Bl/6 J mice cultured in the presence of CSF-1 were treated with Formoterol (10 ng/mL, cat # 1448, Tocris) or a vehicle for 1 h. The supernatant was quickly collected, frozen and kept at −80 °C. Collected supernatants were used to analyze the released inflammatory mediators using the Mouse Inflammation Antibody Array, Membrane 40 Targets (cat # ab133999, Abcam, Berlin, Germany) following the manufacturer’s instruction.
3
Detailed Protocol for Cell and In Vivo Assays
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The following chemicals and reagents were used: 10 μM CL-316243 (MilliporeSigma, C5976) in cells unless otherwise specified and 0.1 mg/kg in mice, 17 ng/mL TNF-α (MilliporeSigma, T7539-50 μg), 1–10 μM ESI-09 (cell culture APExBIO, B4814-5.1), 10 mg/kg ESI-09 (in vivo, MilliporeSigma, 5.00506.0001), Lipofectamine RNAiMAX (Life Technologies, 13778030), 5 μM U73122 (Tocris, 1268), 5 μg/mL cycloheximide (MilliporeSigma, 239765), 50 μM BAPTA-AM (Fisher, 50-136-4911), 4 μM IKK16 (Fisher, IKK INHIBITOR VII/10MG NC1291993), 0.005–50 μM FSK (SellekChem, S2449), Fura2-AM (MilliporeSigma, F0888-5MG), 0.001–10 μM dobutamine (Tocris, 051550), and 0.0001–10 μM formoterol (Tocris, 144810).
4
Characterizing cAMP Signaling in HEK293G Cells
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The cAMP GloSensor™ Human Embryonic Kidney 293 (HEK293G) cell line and GloSensor™ cAMP reagent were purchased from Promega (Madison, WI, USA). Isoprenaline hydrochloride, salmeterol, ICI‐118551 , propranolol , bisoprolol , 3‐isobutyl‐1‐methylxanthine (IBMX), rolipram, Dulbecco's Modified Eagle Medium (DMEM), L‐glutamine, phosphate buffered saline (PBS), trypsin–EDTA, foetal calf serum (FCS) and poly‐D‐lysine were all obtained from Sigma‐Aldrich (St. Louis, MO, USA). Forskolin, formoterol and salbutamol hemisulfate were obtained from Tocris Bioscience (Bristol, UK). Carvedilol was obtained from ACROS Organics (Geel, Belgium). Any other chemicals used were from Sigma‐Aldrich (St. Louis, MO, USA).
5
Formoterol Administration in Mice
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Mice were i.p. injected with 50 μg/kg of Formoterol (cat # 1448, Tocris, Wiesbaden-Nordenstadt, Germany) [32 (link)] or its solvent (0.9% NaCl) 1 h before behavioral analysis or perfusion for immunofluorescence experiments.
6
Characterization of Cardiac Cell Lines
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Streptavidin paramagnetic beads were purchased from Promega (Wisconsin, USA). Blasticidin, StemPro34, Activin A, RPMI, GlutaMAX, B27, secondary antibodies, Essential 8 Medium, and the Nunc Lab-tek chambered coverglass (155411) were purchased from Thermo Fisher Scientific (Paisley, UK). BMP4 and anti-CD90 were purchased from R&D systems (Minneapolis, USA). DMEM, FBS, PBS, NEAA, trypsin, poly-D-lysine, CGP 12177, CGP 20712A, ICI 118551, propranolol, salmeterol, salbutamol, Hoechst 33342, DAPI, anti α-actinin, and formalin were purchased from Sigma-Aldrich (Missouri, USA). bFGF was purchased from PeproTech (London, UK). Matrigel (#356235) was purchased from Corning (New York, USA). Black and white 96-well plates were purchased from Greiner bio-one (Kremsmunster, Austria). Formoterol and isoprenaline were purchased from Tocris Bioscience (Bristol, UK). 3 H-CGP 12177 and MicroScint 20 were purchased from PerkinElmer (Massachusetts, USA). SNAP-Surface Alexa Fluor 488 was purchased from New England Biolabs (Hitchin, UK). All other chemicals were purchased from Sigma-Aldrich.
The HUES7 cell line was a kind gift by Chad Cowan and Doug Melton. 51 The BL15 was provided by David Darling. 56 The HEK293T cell line was obtained from the American Type Culture Collection (CRL-11268).
The HUES7 cell line was a kind gift by Chad Cowan and Doug Melton. 51 The BL15 was provided by David Darling. 56 The HEK293T cell line was obtained from the American Type Culture Collection (CRL-11268).
7
Leukocyte Activation Pathway Assay
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Epinephrine, norEpinephrine, Escherichia coli 0111:B4 lipopolysaccharide (LPS) and Accutase ® were purchased from Sigma-Aldrich (St Quentin-Fallavier, France). Formoterol (b2-AR agonist) and were obtained from Tocris Biosciences (Bristol, UK) and Pam2CSK4 originated from InvivoGen (Toulouse, France). Recombinant human Macrophage-Colony Factor (rhM-CSF) was purchased from Miltenyi-Biotec (Bergisch Gladbach, Germany) and Alexa Fluor ® 647 succinimidyl ester (AF647) fluorescent dye came from Thermo Fisher Scientific (Paysley, UK).
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