Cgp 20712a

Manufactured by Bio-Techne

Sourced in United Kingdom

The CGP 20712A is a compact and versatile laboratory equipment designed for a variety of applications. It functions as a powerful precision balance, capable of accurately measuring the weight of samples with high precision. The device features a digital display and intuitive controls for easy operation.

Automatically generated - may contain errors

Lab products found in correlation

Ici 118 551, by Bio-Techne (4 mentions) Propranolol, by Bio-Techne (2 mentions) Cimaterol, by Bio-Techne (2 mentions) Cgp 12177, by Bio-Techne (2 mentions) Bodipy tmr cgp cgp 12177 tmr, by Thermo Fisher Scientific (2 mentions) 3h cgp12177, by PerkinElmer (2 mentions) Isoprenaline, by Merck Group (1 mentions) Nanoluc substrate furimazine, by Promega (1 mentions) Snap surface alexa fluor 488, by New England Biolabs (1 mentions) Cell culture reagents, by Merck Group (1 mentions) Bodipy tmr cgp, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by GE Healthcare (1 mentions) Furimazine, by Promega (1 mentions) Microscint 20, by PerkinElmer (1 mentions) Decyl β d maltopyranoside, by Anatrace (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (1 mentions) 14c camp, by PerkinElmer (1 mentions) 3h adenine, by PerkinElmer (1 mentions) Fugene hd transfection reagent, by Promega (1 mentions) Ultima gold scintillation fluid, by PerkinElmer (1 mentions)

6 protocols using cgp 20712a

Fluorescent Ligands for GPCR Studies

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propranolol‐Peg8‐BY630 (propranolol‐peg8‐BODIPY630/650) and prop‐β(Ala‐Ala)‐BY630 (propranolol‐βalanine‐ βalanine‐X‐BODIPY630/650) were obtained from CellAura (Nottingham, UK). BODIPY‐TMR‐CGP (CGP‐12177‐TMR) was purchased from Molecular Probes (Eugene, OR). propranolol, ICI 118551, CGP 12177, CGP 20712A, and cimaterol were from Tocris (Bristol, UK). Isoprenaline was purchased from Sigma‐Aldrich (Gillingham, UK). The NanoLuc substrate furimazine was obtained from Promega (Southampton, UK). The radioligand ³H CGP 12177 was obtained from PerkinElmer (Coventry, UK).

Soave M., Stoddart L.A., Brown A., Woolard J, & Hill S.J. (2016). Use of a new proximity assay (NanoBRET) to investigate the ligand‐binding characteristics of three fluorescent ligands to the human β 1‐adrenoceptor expressed in HEK‐293 cells. Pharmacology Research & Perspectives, 4(5), e00250.

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Fluorescent Ligand Binding Assay

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Cell culture plastics were purchased from Fisher Scientific (Loughborough, UK) and cell culture reagents were from Sigma Aldrich (Gillingham, UK) except for fetal calf serum, which was obtained from PAA Laboratories (Pasching, Austria). SNAP-Surface™ Alexa Fluor ®488 was obtained from New England Biolabs (Ipswich, MA, USA). BODIPY-TMR-CGP was from Molecular Probes (Leiden, the Netherlands) and unlabelled CGP 12177, propranolol and CGP 20712A were from Tocris Cookson (Avonmouth, Bristol, UK). All other reagents were from Sigma Chemicals (Poole, UK).

Gherbi K., Briddon S.J, & Hill S.J. (2014). Detection of the secondary, low-affinity β1-adrenoceptor site in living cells using the fluorescent CGP 12177 derivative BODIPY-TMR-CGP. British Journal of Pharmacology, 171(23), 5431-5445.

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Radioligand Binding Assay for β1-Adrenoceptor

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[³H]-adenine, [³H]-CGP 12177, [¹⁴C]-cAMP, Microscint-20 and Ultima Gold Scintillation fluid were from Perkin Elmer (Coventry, West Midlands, UK). CGP 12177, CGP 20712A, cimaterol and ICI 118551 were from Tocris Bioscience (Bristol, UK). Decyl-β-D-maltopyranoside was from Anatrace (Berkshire, UK). Purified β₁-m23-StaR [8] (link) was provided by Heptares Therapeutics. The following antibodies were used: goat-anti-mouse-Rhodamine secondary antibody (Molecular Probes, Life Technologies, Paisley, UK), horse radish peroxidase-conjugated secondary antibody (Cell Signalling Technology, Leiden, The Netherlands). BODIPY-TMR-CGP (CGP-12177-TMR) was purchased from Molecular Probes (Eugene, OR, USA). Fugene HD transfection reagent and furimazine were from Promega (Southampton, UK). Purified turkey and human ECL2 peptides were obtained from Cambridge Research Biochemicals (Cambridge, UK). All other reagents were from Sigma-Aldrich (Gillingham, UK).

Soave M., Cseke G., Hutchings C.J., Brown A.J., Woolard J, & Hill S.J. (2018). A monoclonal antibody raised against a thermo-stabilised β1-adrenoceptor interacts with extracellular loop 2 and acts as a negative allosteric modulator of a sub-set of β1-adrenoceptors expressed in stable cell lines. Biochemical Pharmacology, 147, 38-54.

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Pharmacological Inhibitors of Cell Signaling

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Erythro-9-(2-hydroxy-3-nonyl) adenine, cilostamide, rolipram, MDL 12330A, CGP 20712A, and ICI 118,551 were purchased from Tocris Bioscience (Bristol, UK). MEM, penicillin-streptomycin, and insulin-transferrin-selenium were purchased from Life Technologies (Carlsbad, CA). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO).

Rudokas M.W., Post J.P., Sataray-Rodriguez A., Sherpa R.T., Moshal K.S., Agarwal S.R, & Harvey R.D. (2021). Compartmentation of β2-Adrenergic Receptor Stimulated cAMP Responses by Phosphodiesterases Type 2 and 3 in Cardiac Ventricular Myocytes. British journal of pharmacology, 178(7), 1574-1587.

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Modulation of Microglial Inflammatory Response

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Mixed glial cells were obtained from the cerebral cortex of Sprague Dawley rat pups at postnatal days 1–3 and cultured in DMEM supplemented with 10% fetal bovine serum and 1% Penicillin/Streptomycin. After 10 days in vitro, microglia were harvested by gentle shaking of the growth flask, plated in a 96 well plate at a density of 30,000 cells/well, and incubated at 37°C overnight. Microglia were pretreated with the ADRB1 antagonists CGP 20712A (0.1 μM; Tocris 1024) or betaxolol (10 μM; Tocris 0906), the ADRB2 antagonist ICI-118551 (0.1 μM; Tocris 0821), or vehicle for 15 min. Following the pretreatment, microglia were stimulated with LPS (10 ng/ml; Sigma Aldrich L4391) along with or without xamoterol (S-enantiomer; 1 μM) or isoproterenol (1 μM; Sigma Aldrich I5627) for 4 h at 37°C. Following the 4 h incubation, cell media was collected and concentration of TNF-α was measured by ELISA (Invitrogen, KRC3011) according to the manufacturer’s instruction.

Ardestani P.M., Evans A.K., Yi B., Nguyen T., Coutellier L, & Shamloo M. (2017). Modulation of neuroinflammation and pathology in the 5XFAD mouse model of Alzheimer’s Disease using a biased and selective beta-1 adrenergic receptor partial agonist. Neuropharmacology, 116, 371-386.

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Modulating β-Adrenergic Receptor Signaling

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β1-AR stimulation was achieved with 10 and 100 nM isoproterenol in the presence of the β2 antagonist ICI 118,551 (ICI; 100 nM). Selective β2-AR stimulation was achieved with 50 and 100 nM zinterol (Tocris) in the presence of the β1 antagonist CGP20712A (CGP; 10 nM). Myocyte populations used to measure phospho-protein responses were field-stimulated during β-AR stimulation. At 5 min, perfusion buffer was rapidly aspirated and replaced with Laemmli sample buffer containing protease inhibitor cocktail (Roche) and phosphatase inhibitor cocktail (Thermo Fisher Scientific).
For experiments using pertussis toxin (PTX) to disable Gαi, cells were treated with 1.5 µg.ml⁻¹ PTX for 3 h at 37°C. To determine the contribution of HMG CoA reductase inhibition and depletion of isoprenoids to basal and β-AR responses, mevalonate (100 µM), FPP and/or GGPP (10 µM) were added to statin-containing medium at the start of the 48 h culture period. For these protocols, shortening was measured in the absence of fura-loading.

Pugh S.D., MacDougall D.A., Agarwal S.R., Harvey R.D., Porter K.E, & Calaghan S. (2014). Caveolin Contributes to the Modulation of Basal and β-Adrenoceptor Stimulated Function of the Adult Rat Ventricular Myocyte by Simvastatin: A Novel Pleiotropic Effect. PLoS ONE, 9(9), e106905.

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