Diazepam

Manufactured by Pfizer

Sourced in United States

Diazepam is a laboratory equipment product used for various analytical and research purposes. It serves as a reference standard for the identification and quantification of diazepam, a benzodiazepine drug, in samples. The core function of Diazepam is to provide a reliable and consistent reference material for analytical procedures.

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Lab products found in correlation

Pilocarpine hydrochloride, by Merck Group (6 mentions) Diazepam, by Merck Group (3 mentions) Lithium chloride (licl), by Merck Group (3 mentions) Methylscopolamine, by Merck Group (3 mentions) L4516 1mg, by Merck Group (2 mentions) Pilocarpine, by Merck Group (2 mentions) Potassium dihydrogen phosphate, by Merck Group (2 mentions) Potassium hydroxide, by Merck Group (2 mentions) Dipotassium hydrogen phosphate, by Merck Group (2 mentions) Pyrogallol solution, by Merck Group (2 mentions) Alfaxalone, by Merck Group (1 mentions) Alfaxalone, by Pfizer (1 mentions) Propofol, by Pfizer (1 mentions) Dl apv, by Merck Group (1 mentions) Propofol, by MP Biomedicals (1 mentions) Kainic acid, by Cayman Chemical (1 mentions) Methylatropine bromide, by Merck Group (1 mentions) Lactic acid, by Merck Group (1 mentions) Pyridine 2 aldoxime methochloride 2 pam, by Merck Group (1 mentions) Paraoxon, by Chem Service (1 mentions)

29 protocols using diazepam

Electrophysiology and Behavioral Experiments with Anesthetic Agents

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The salts used in buffers, alfaxalone, diazepam, CNQX and DL-APV used in electrophysiological experiments were bought from Sigma-Aldrich (St. Louis, MO). Propofol was purchased from MP Biomedicals (Solon, OH). Stock solution of alfaxalone was made in DMSO at 10 mM and stored at room temperature. Stock solution of Propofol was made in DMSO at 200 mM and stored at room temperature. For electrophysiological recordings, stock solution of diazepam was made in ND96 at 30 μM and stored at −20 °C. Dilutions to lower concentrations were made from that solution. The highest final concentration of DMSO was 0.3% (v/v). This concentration of DMSO has been shown to be without effect on currents from recombinant α1β2γ2L GABA_A receptors or GABA_A receptor-mediated synaptic currents^{52 (link),53 (link)}.
Pharmaceutical grade reagents were used in all behavioral experiments. Propofol (10 mg/ml, Hospira, Inc., San Jose, CA), alfaxalone (10 mg/ml, Jurox, Inc., Kansas City, MO), diazepam (5 mg/ml, Hospira, Inc.), combinations of either Propofol and alfaxalone, or diazepam and alfaxalone, or sterile normal saline (20 Units) were injected intraperitoneally using U-100 insulin syringes. diazepam was diluted to 0.5 mg/ml with normal saline prior to injection.

Cao L.Q., Montana M.C., Germann A.L., Shin D.J., Chakrabarti S., Mennerick S., Yuede C.M., Wozniak D.F., Evers A.S, & Akk G. (2018). Enhanced GABAergic actions resulting from the coapplication of the steroid 3α-hydroxy-5α-pregnane-11,20-dione (alfaxalone) with propofol or diazepam. Scientific Reports, 8, 10341.

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Kainic Acid-Induced Epilepsy Model

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One week after surgery, kainic acid (5.3 mg/ml PBS; Cayman Chemical, Ann Arbor, MI, USA) was injected intraperitoneally at a dose of 25–100 mg kainic acid/kg mouse body weight (in increments of 10–25 mg/kg) until the mouse went into status epilepticus, defined as persistent epileptiform activity on the EEG accompanied by recurrent clinical seizures for at least 12 hours (the minimum time we determined in pilot experiments to induce epilepsy). Some mice received diazepam i.p. (5 mg/kg) (Hospira, Lake Forest, IL, USA) if clinically evident convulsions were persistent and severe. Mice were injected with PBS (i.p.) 4–8 hr after induction of status epilepticus to maintain adequate hydration.

Holden K, & Hartman A.L. (2017). D-leucine: evaluation in an epilepsy model. Epilepsy & behavior : E&B, 78, 202-209.

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Pilocarpine-Induced Epilepsy Model in Rats

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These experiments utilized the method described in Luna-Munguia et al. (2017) (link). Briefly, 56-days-old rats were randomly assigned to a group that received an injection of pilocarpine hydrochloride (340 mg/kg ip; Sigma-Aldrich, St. Louis, MO) 20 min after a single intraperitoneal injection of methylatropine bromide (5 mg/kg; Sigma-Aldrich, St. Louis, MO). Five to ten minutes after the pilocarpine administration, the animals started with head nodding, evolving into recurrent generalized convulsions (status epilepticus) within 40 min. Rats that did not develop status epilepticus within this period of time, received an additional dose of pilocarpine hydrochloride (170 mg/kg; ip). Animals were behaviorally monitored by experienced researchers and after 90 min of status epilepticus, seizures were interrupted with diazepam (10 mg/kg ip; Hospira, Lake Forest, IL). Control animals received a single injection of methylatropine bromide (5 mg/kg; ip) 20 min prior to vehicle (0.9% saline solution; ip), and treated with diazepam 2 h after the saline injection. Monitoring of the spontaneous seizures began immediately after the pilocarpine injection and was continued for 14 days using a video recording system. Rats with epilepsy that did not achieve the initial body weight within the first 12 days after pilocarpine-induced status epilepticus were excluded from the study.

Luna-Munguia H., Zestos A.G., Gliske S.V., Kennedy R.T, & Stacey W.C. (2018). Chemical biomarkers of epileptogenesis and ictogenesis in experimental epilepsy. Neurobiology of disease, 121, 177-186.

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Chemogenetic Modulation of Neural Circuits

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CNO (Enzo life science-Biomol, BML-NS105-0005) were freshly dissolved in injection saline (0.9% NaCl) and intraperitoneally injected at 5 mg/kg for hM4Di silencing, or 2 mg/kg for hM3Dq activation. Other compounds used for intraperitoneal injection: CCK (5 μg/kg, Tocris), Diazepam (1 mg/kg or 2 mg/kg, Hospira), LiCl (150 mg/kg, Sigma, prepared in 150 mM), LPS (0.1 mg/kg, Sigma, L4516-1MG). Behavioral test were usually performed 20-30 min after drug injection.

Cai H., Haubensak W., Anthony T, & Anderson D.J. (2014). Central amygdala PKC-δ+ neurons mediate the influence of multiple anorexigenic signals. Nature neuroscience, 17(9), 1240-1248.

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Pharmacological Evaluation of Analgesics

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Fentanyl HCl, morphine sulfate, buprenorphine HCl, naltrexone HCl, U69593, psilocybin, and amphetamine sulfate were provided by the National Institute on Drug Abuse Drug Supply Program and were dissolved in sterile saline. Nalbuphine HCl (provided by Dr. Kenner Rice, National Institute on Drug Abuse Intramural Research Program) was also dissolved in sterile saline. NAQ {17-Cyclopropyl-methyl-3,14β-dihydroxy-4,5α-epoxy-6α-[(3′-isoquinolyl)acetamido]-morphinan; provided by Dr. Yan Zhang, Virginia Commonwealth University} was dissolved in 10% DMSO and 90% water. Ketoprofen (100 mg/ml; Ford Dodge, IA) was diluted in sterile saline. Diazepam (5 mg/ml, Hospira, Lake Forest, IL) was diluted in 1:4:5 ethanol, propylene glycol, and saline. All drugs were administered subcutaneously (SC) in volumes of 10 ml/kg. Lactic acid and lithium chloride were purchased from Sigma-Aldrich (St. Louis, MO). Lactic acid was diluted in sterile water, while lithium chloride was dissolved in saline. Both were administered intraperitoneally (IP) in a volume of 10 ml/kg.

Negus S.S., Akbarali H.I., Kang M., Lee Y.K., Marsh S.A., Santos E.J, & Zhang Y. (2023). Role of mu opioid receptor (MOR) agonist efficacy as a determinant of opioid antinociception in a novel assay of pain-depressed behavior in female and male mice. Frontiers in Pain Research, 4, 1281698.

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Pharmacological Reagent Preparation Protocol

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Paraoxon was purchased from Chem Service (West Chester, PA). Pilocarpine hydrochloride and pyridine-2-aldoxime methochloride (2-PAM) were purchased from Sigma Aldrich (St. Louis, MO). Diazepam (Hospira, Lake Forest, IL) was purchased as a stock solution dissolved in 0.9% saline from the University of Washington Medical Center Pharmacy. SR141716 was obtained from the NIDA Drug Supply Program (Bethesda, MD) and was prepared in pharmasolve/cremophor RH40 (pharmasolve: cremophor RH40: drug, 1:9:40). CP55940 was obtained from the NIDA Drug Supply Program and was prepared in a vehicle solution consisting of cremophor RH40: ethanol: saline (1:1:18). All drugs except for SR141716 and CP55940 were made as stock solutions in 0.9% saline.

Kow R.L., Cheng E.M., Jiang K., Le J.H., Stella N, & Nathanson N.M. (2014). Muscarinic M1 receptor and cannabinoid CB1 receptor do not modulate paraoxon-induced seizures. Pharmacology Research & Perspectives, 3(1), e00100.

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Preparation of Psychoactive Compounds

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Pilocarpine hydrochloride was purchased from Sigma Aldrich and dissolved in 0.9% saline. Diazepam (Hospira) and phenobarbitol (West-Ward) were purchased as stock solutions dissolved in 0.9% saline from the University of Washington Medical Center Pharmacy. SR141716 was obtained from the NIDA Drug Supply Program and was prepared in pharmasolve/cremophor RH40 (pharmasolve: cremophor RH40: drug, 1∶9:40). CP55940 was obtained from the NIDA Drug Supply Program and was prepared in a vehicle solution consisting of cremophor RH40: ethanol: saline (1∶1:18).

Kow R.L., Jiang K., Naydenov A.V., Le J.H., Stella N, & Nathanson N.M. (2014). Modulation of Pilocarpine-Induced Seizures by Cannabinoid Receptor 1. PLoS ONE, 9(4), e95922.

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Pharmacological Management of Seizure Disorders in Rats

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As described previously,^{7 (link)} rats received lithium chloride (5 mEq/kg; #L-0505 Sigma, St. Louis MO, USA) subcutaneously and, 16 hours later, i.p. pilocarpine hydrochloride (320 mg/kg; #P6503 Sigma) and scopolamine methyl bromide (1mg/kg; i.p., #S8502; Sigma). Seizures started 7.6 ± 2.7 min. after pilocarpine injection, and the second stage 3 or higher seizure occurred 8–20 min. later. At the end of the second stage 3 or higher seizure, all animals received scopolamine (10 mg/kg i.p.; #S1013; Sigma) to remove the original seizure trigger without stopping SE, and either sham injection (SE control group), one drug, or a combination of 3 drugs i.p.. Drugs included diazepam (#321312 Hospira), ketamine (#RL3760 Hospira) and sodium valproate (#P4543 Sigma).

Niquet J., Baldwin R., Suchomelova L., Lumley L., Eavey R, & Wasterlain C.G. (2017). Treatment of experimental status epilepticus with synergistic drug combinations. Epilepsia, 58(4), e49-e53.

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Pilocarpine-Induced Rat Seizure Model

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To minimize the peripheral effects of pilocarpine, rats were injected intraperitoneally with scopolamine (1 mg/kg) 30 minutes before administration of pilocarpine (385 mg/kg). According to a standard protocol, if rats did not exhibit convulsive seizures within 1 h of pilocarpine injection a maximum of two subsequent doses of pilocarpine (192.5 mg/kg) were given in order to produce equivalence in seizures between animals (Brooks-Kayal et al., 1998 (link); Shumate et al., 1998). Diazepam (6 mg/kg; Hospira, Lake Forest, IL) was administered 1 h after the onset of SE in order to stop seizure progression and additional doses (3 mg/kg) were administered every 2 h until rats stopped seizing. Control rats were handled similarly but treated with a subconvulsive dose of pilocarpine (38.5 mg/kg) and 1/10 of the dose of Diazepam (0.6 mg/kg).

González M.I., Grabenstatter H.L., del Rio C.C., Del Angel Y.C., Carlsen J., Laoprasert R., White A.M., Huntsman M.M, & Brooks-Kayal A. (2015). Seizure-Related Regulation of GABAA Receptors in Spontaneously Epileptic Rats. Neurobiology of disease, 77, 246-256.

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D-Leucine Inhibits Kainic Acid-Induced Seizures

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A single dose of each amino acid dissolved in H2O (Sigma-Aldrich, St. Louis, MO, U.S.A) was injected intraperitoneally (i.p.) into 5 week-old mice 3 h before kainic acid injection. The reported purity of the lots of L- and D-leucine used was 99% (based on optical rotation data); L-leucine had no more than 5% of the D-isomer, and D-leucine had 0.4% of the L-isomer (Sigma Aldrich, technical data). In separate experiments, L- and D-leucine was administered at the indicated time (15 or 20 min) after kainic acid injection. The 15–20 min time is midrange for the clinical operational definition of status epilepticus (defined as 5–30 min of seizure activity without a return to baseline, (Lowenstein, 1999 (link))). In these experiments, all mice had experienced at least two seizures with a score ≥ 3 when D-leucine was administered, with seizures starting 10–15 min after kainic acid injection. Diazepam (5 mg/ml injectable solution, diluted 1:1.5 in phosphate-buffered saline; Hospira, Lake Forest, IL, U.S.A.) was injected i.p. at 20 min after kainic acid injection, when all mice had exhibited two or three stage ≥3 seizures.

Hartman A.L., Santos P., O’Riordan K.J., Stafstrom C.E, & Hardwick J.M. (2015). Potent anti-seizure effects of D-leucine. Neurobiology of disease, 82, 46-53.

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