Clonidine

Manufactured by Merck Group

Sourced in United States, Germany, Japan, United Kingdom

Clonidine is a laboratory equipment product used for scientific research and analysis. It is a chemical compound that serves as a central alpha-2 adrenergic receptor agonist. The core function of Clonidine is to induce specific physiological responses in experimental models or cell-based assays, as required by research protocols. A detailed, unbiased, and factual description of Clonidine's intended use or applications is not available.

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Close spelling variants of this product

Clonidine hydrochloride (34 citations) Clonidine HCl (6 citations)

49 protocols using clonidine

Osteoclast Differentiation Assay with Pharmacological Modulators

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Mouse bone marrow cells were plated at densities of 1.2×10⁵ and 1.0×10⁶ cells into 12-well and 60-mm dishes, respectively, and cultured with 10 ng/ml macrophage colony-stimulating factor (M-CSF; PeproTech, Inc., Rocky Hill, NJ, USA) at 37°C for 3 days. The surface-attached cells were used as osteoclast precursors (22 (link)). These cells were cultured with 10 ng/ml M-CSF and 50 ng/ml RANKL (PeproTech, Inc.). A total of 5–20 µM guanabenz (R&D Systems, Inc., Minneapolis, MN, USA) or 10–20 µM xylazine (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) was applied at the same time point as RANKL; 10–20 µM clonidine (Sigma-Aldrich; Merck KGaA) was administered with RANKL or 1 day after RANKL administration. After a 60-h treatment with RANKL at 37°C, cells were fixed in 10% formalin neutral buffer solution at room temperature and stained with TRAP for 1 h at 37°C. The number of TRAP-positive cells containing three or more nuclei was determined. All positive cells in each well were counted using a light microscope (magnification, ×100; Zeiss AG, Oberkochen, Germany).
RAW264.7 cells were plated at 1.0×10⁵ cells into 60-mm dishes and cultured with 25 ng/ml RANKL in the presence or absence of 5–20 µM guanabenz, 10–20 µM clonidine or 10–20 µM xylazine with or without 10–20 µM yohimbine or 10–20 µM idazoxan (Sigma-Aldrich; Merck KGaA) at 37°C for 2–4 days for qPCR analysis.

Hamajima K., Hamamura K., Chen A., Yokota H., Mori H., Yo S., Kondo H., Tanaka K., Ishizuka K., Kodama D., Hirai T., Miyazawa K., Goto S, & Togari A. (2018). Suppression of osteoclastogenesis via α2-adrenergic receptors. Biomedical Reports, 8(5), 407-416.

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Antihypertensive Drugs Modulate Nischarin Expression

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Considering that Nischarin is a target of antihypertensive drugs, we investigated whether such drugs could affect NISCH expression and the abnormal murine behaviors caused by the dysregulation of this gene. About 50,000 U251 cells were plated into 24-well plates, and were treated with either DMSO or 0.1 mM clonidine (#C7897, Sigma-Aldrich) or 0.1 mM tizanidine (#T6950, Sigma-Aldrich) on the second day for 72 h. Total RNA was then harvested, reverse-transcribed into cDNA, and the expression level of NISCH was determined by RT-qPCR. The mice were fed with water containing either DMSO or 0.1 mM clonidine for 72 h before the behavioral experiments.

Yang Z.H., Cai X., Ding Z.L., Li W., Zhang C.Y., Huo J.H., Zhang Y., Wang L., Zhang L.M., Li S.W., Li M., Zhang C., Chang H, & Xiao X. (2023). Identification of a psychiatric risk gene NISCH at 3p21.1 GWAS locus mediating dendritic spine morphogenesis and cognitive function. BMC Medicine, 21, 254.

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Cardiovascular Tissue Preparation

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ATP, Clonidine, GM, N-Acetyl-L-cysteine (ACC), Tempol all from Sigma (Deisenhofen, Germany), Zym (InvivoGen, San Diego, USA), Milrione (Hikma, Portugal). 2-Deoxyadenosine monohydrate (2-DAM), CPG 20712, ICI 118,551 all from Tocris (Bristol, UK). Epinephrine (INFECTOPHARM GmbH, Heppenheim, Germany).
HEPES solution: 20 mM HEPES, 4.5 mM KCl, 2.5 mM CaCl₂, 11 mM Glucose, 140 mM NaCl, 1 mM MgCl₂, pH was adjusted to 7.4 at 30°C or 4–8°C for tissue preparation using NaOH (4 M).

Weiterer S., Kohlen T., Veit F., Sachs L., Uhle F., Lichtenstern C., Weigand M.A, & Henrich M. (2015). Galactomannan and Zymosan Block the Epinephrine-Induced Particle Transport in Tracheal Epithelium. PLoS ONE, 10(11), e0143163.

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Adrenergic and Corticosteroid Receptor Assay

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For testing adrenergic receptor specificity, seven days after establishment of a latent infection, media containing acyclovir was removed and Complete Neuro media with no acyclovir and with one, two, or three adrenergic agonists were added. Adrenergic receptor (AR) agonists included phenylephrine (α-1 AR agonist), clonidine (α-2 AR agonist), dobutamine (β-1 AR agonist), or terbutaline (β-2 AR agonist) (Sigma). Adrenergic receptor antagonists phentolamine (nonspecific α-AR antagonist) or timolol (nonspecific β-AR antagonist) were also added with 10 µM epinephrine to fresh Complete Neuro media. For testing corticosterone receptor specificity, Complete Neuro media with no acyclovir and with either the glucocorticoid receptor agonist dexamethasone (GR agonist) or mineralocorticoid receptor agonist aldosterone (MR agonist) (Sigma) were added to Complete Neuro media (10 µM). The corticosterone antagonists mifepristone (GR antagonist), eplerenone (MR antagonist) (Sigma), or both at 10 µM were added to fresh Complete Neuro media with 10 µM corticosterone-HBC.

Goswami P., Ives A.M., Abbott A.R, & Bertke A.S. (2022). Stress Hormones Epinephrine and Corticosterone Selectively Reactivate HSV-1 and HSV-2 in Sympathetic and Sensory Neurons. Viruses, 14(5), 1115.

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Clonidine Perfusion Bath Preparation

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A stock solution of clonidine (Sigma-Aldrich, St. Louis, MO) was prepared in de-ionized water at a stock concentration of 10 mM and diluted on the day of the experiment to a final concentration in the perfusion bath of 10 μM.

Reyes B.A., Carvalho A.F., Szot P., Kalamarides D.J., Wang Q., Kirby L.G, & Van Bockstaele E.J. (2017). Cortical adrenoceptor expression, function and adaptation under conditions of cannabinoid receptor deletion. Experimental neurology, 292, 179-192.

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Oligonucleotide and Small Molecule Protocols

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All oligonucleotide sequences and modification patterns synthesized in-house are summarized in Supplementary Table S1. Accell siRNA (Thermo Fisher) targeting HTT was purchased and used without further purification. Clonidine (Sigma–Aldrich), Retro-1 (Sigma–Aldrich) and Salubrinal (Sigma–Aldrich) were dissolved in DMSO and stored as 10 mM stock solutions at −20°C. Unless otherwise specified, all oligonucleotides and small molecules were diluted in OptiMEM (Gibco) prior to administration in vitro.

Osborn M.F., Alterman J.F., Nikan M., Cao H., Didiot M.C., Hassler M.R., Coles A.H, & Khvorova A. (2015). Guanabenz (Wytensin™) selectively enhances uptake and efficacy of hydrophobically modified siRNAs. Nucleic Acids Research, 43(18), 8664-8672.

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Hypoxia-Induced Microglial Cell Stress

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The BV2 cells (mouse microglial cells) were purchased from ATCC Company (Milan, Italy). The cells were suspended in DMEM (Dulbecco’s Modified Eagle Medium containing 10% fetal bovine serum (FBS), 100 U/mL of penicillin, and 100 U/mL of streptomycin) culture medium. At 80% confluency, the cells were treated with trypsin-EDTA solution (0.05% trypsin and 0.02% EDTA), and the resulting cell suspension was adjusted to a cell density of 5 × 104 cells/mL. The cell suspensions were washed with PBS (Phosphate-Buffered Saline), resuspended in serum-free DMEM medium, and placed in an incubator at 37 °C, without and with the addition of Propofol, Dexmedetomidine, or Clonidine (Sigma–Aldrich, Milan, Italy). The drugs were tested at final concentrations of 25 and 50 μM. Hypoxia was then induced by challenging the cell cultures with a gas mixture containing 1.0% O2, 5% CO2, and 37 °C for 3 h to initiate hypoxia, followed by 3 h of reoxygenation at 37 °C using a gas mixture containing 5% CO2 and 18.0% O2. We used gas-controlled incubators to control the O2 levels of the cell cultures. In hypoxia, the O2 levels were 1%, and the CO2 levels were 5%. Under the reoxygenation conditions, the O2 levels were 18%, and the CO2 levels were 5%. Nitrogen was added to the incubator in order until the set O2% was achieved.

Longhitano L., Distefano A., Murabito P., Astuto M., Nicolosi A., Buscema G., Sanfilippo F., Lazzarino G., Amorini A.M., Bruni A., Garofalo E., Tibullo D, & Volti G.L. (2022). Propofol and α2-Agonists Attenuate Microglia Activation and Restore Mitochondrial Function in an In Vitro Model of Microglia Hypoxia/Reoxygenation. Antioxidants, 11(9), 1682.

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Microinfusion of Receptor Antagonists

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For microinfusions, the concentrations used were 21.1 mM (6.25 μg μl⁻¹) for β-adrenoceptor antagonist propranolol [(S)-(−)-propranolol hydrochloride, 295.80 g mol⁻¹; Sigma-Aldrich), 3.1 mM (1 μg μl⁻¹) for D₁/D₅ receptor antagonist SCH23390 (SCH 23390 hydrochloride, 324.24 g mol⁻¹; Tocris) and 2% w/v for voltage-gated sodium channel blocker lidocaine (lidocaine hydrochloride monohydrate, 288.81 g mol⁻¹; Sigma-Aldrich). α₂-adrenoceptor agonist clonidine (clonidine hydrochloride, 266.55 g mol⁻¹; Sigma-Aldrich) was administered i.p. at a dose of 50 μg kg⁻¹ of body weight. We used 0.9% NaCl (saline) in H₂O as a vehicle and for control infusions. Both vehicle and drug solutions were stored in 100 μl aliquots at −20°C until use.

Takeuchi T., Duszkiewicz A.J., Sonneborn A., Spooner P.A., Yamasaki M., Watanabe M., Smith C.C., Fernández G., Deisseroth K., Greene R.W, & Morris R.G. (2016). Locus coeruleus and dopaminergic consolidation of everyday memory. Nature, 537(7620), 357-362.

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Clonidine Treatment in Hypertensive Rats

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SHR aged between 12 and 14 weeks were treated with 0.1 mg·kg^-1·day^-1 clonidine (Sigma-Aldrich, USA) dissolved in distilled water once per day for 28 days by gavage. The clonidine dose was chosen according to previous studies from our laboratory, showing that these doses were adequate to inhibit central sympathetic activity and also to normalize arterial pressure in hypertensive animals (19 (link)). The treatment was performed by the introduction of the solution via a stainless-steel gavage needle (cannula diameter, 1.2 mm) attached to a syringe, and all procedures were performed at the same time of day. The normotensive control group consisted of WKY rats treated with vehicle (WKY+VEH) and SHR were divided into two groups of animals (minimum of 10 animals per group), treated with vehicle (SHR+VEH) or clonidine (SHR+CLO).

Nisimura L.M., Bousquet P., Muccillo F., Tibirica E, & Garzoni L.R. (2020). Tyrosine hydroxylase and β2-adrenergic receptor expression in leukocytes of spontaneously hypertensive rats: putative peripheral markers of central sympathetic activity. Brazilian Journal of Medical and Biological Research, 53(12), e9615.

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Modulation of Locus Coeruleus Activity

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Both male and female mice were used in this study. All mice were C57BL/6J except 2 (out of 6) included in the localized Clonidine treatment were of mixed B6J/129 background. Mice were housed with reversed light/dark cycle (9A – 9P dark, 9P – 9A light). Mice of 6–12 weeks were implanted with head posts and/or cannulae. Clonidine (an α2 agonist, Sigma-Aldrich) was administered locally in 6 mice and systemically in 3 mice. Yohimbine (an α2 antagonist, Sigma-Aldrich) was administered locally in 7 mice and systemically in 5 mice. Every mouse received corresponding localized or systemic saline injections as controls. Quantification of localized pharmacological effects on LC activity was performed by immunostaining for the immediate early gene c-fos in 11 mice. All procedures were approved by the UC Riverside Animal Care and Use Committee.

McBurney-Lin J., Sun Y., Tortorelli L.S., Anh Nguyen Q., Haga-Yamanaka S, & Yang H. (2020). Bidirectional pharmacological perturbations of the noradrenergic system differentially affect tactile detection. Neuropharmacology, 174, 108151.

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