1 2 chlorophenyl diphenylmethyl 1h pyrazole tram 34

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Sourced in United States, Germany, Australia

1-(2-Chlorophenyl) diphenylmethyl-1H-pyrazole (TRAM-34) is a potent and selective blocker of the intermediate-conductance calcium-activated potassium channel (KCa3.1). It is a commonly used research tool for the study of KCa3.1 channel function.

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TRAM-34 (41 citations) Pyrazole (26 citations)

4 protocols using 1 2 chlorophenyl diphenylmethyl 1h pyrazole tram 34

Vasodilatory Mechanisms in Isolated Arteries

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The following drugs were used: acetylcholine, 4-aminopyridine, apamin, 4′-hydroxy-3′-methoxyacetophenone (apocynin), barium chloride, cocaine, glibenclamide, guanethidine, 5-HT, iberiotoxin, indomethacin, L-NOARG, N^omega-propyl-L-arginine, phentolamine, ouabain, prazosin, propranolol, SOD, tetrodotoxin, TEA and 1-(2-Chlorophenyl) diphenylmethyl- 1H-pyrazole (TRAM-34) (Sigma Chemical Co, St Louis, MO, USA). NS-398, SC-560 and 1400 W were purchased from Tocris Cookson (Bristol, UK). Bosentan was a gift from F. Hoffman-La Roche Laboratories (Basel, Switzerland).
All drugs were added in volumes not exceeding 0.3% of the organ baths to reach the final required concentration. They were dissolved in distilled water with the exception of indomethacin which was prepared in ethanol (96%), glibenclamide, SC-560 and NS-398 which required dimethylsulphoxide (10%). Preliminary experiments revealed no effects of the solvent used on the contractility of the preparations. Stock solutions were prepared and stored at −20°C and fresh solutions were prepared daily.

Martínez A.C., Hernández M., Novella S., Martínez M.P., Pagán R.M., Hermenegildo C., García-Sacristán A., Prieto D, & Benedito S. (2014). Diminished Neurogenic Femoral Artery Vasoconstrictor Response in a Zucker Obese Rat Model: Differential Regulation of NOS and COX Derivatives. PLoS ONE, 9(9), e106372.

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Comprehensive Pharmacological Compound Protocol

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2,6-Bis(2-benzimidazolyl)pyridine (BBP), dofetilide, bicuculline methiodide, paxilline, 1-[(2-chlorophenyl)diphenylmethyl]-1H-pyrazole (TRAM-34) and 4-aminopyridine were purchased from Sigma-Aldrich (Germany). 2-(4-Chlorophenoxy)-2-methyl-N-[5-[(methylsulfonyl)amino]tricyclo[3.3.1.13,7]dec-2-yl]-propanamide (JNJ 303) was purchased from Tocris (United Kingdom) and N-(2-{[(1R)-1-[3-(trifluoromethyl)phenyl]ethyl]amino}-1H-1,3-benzodiazol-4-yl)acetamide (AP14145) was synthetized by Syngene Inc. (Bangalore, India). All compounds were dissolved in pure dimethyl sulfoxide (Sigma-Aldrich, Germany) into 10 mM stock solutions and stored at −20°C. On the day of the experiment, the stock solution was thawed and the corresponding amount of compound was added to the working solution.

Simó-Vicens R., Bomholtz S.H., Sørensen U.S, & Bentzen B.H. (2018). 2,6-Bis(2-Benzimidazolyl)Pyridine (BBP) Is a Potent and Selective Inhibitor of Small Conductance Calcium-Activated Potassium (SK) Channels. Frontiers in Pharmacology, 9, 1409.

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Whole-Cell Patch Clamp Experiments

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For the whole-cell patch clamp experiment, low Ca²⁺-buffered KCl pipette solution contained (in mM): 140 KCl, 10 HEPES, 5 NaCl, 3 Mg-ATP, 0.1 EGTA, 1 MgCl₂ and was adjusted to pH 7.2 with KOH. Ca²⁺-clamped KCl pipette solution contained (in mM) 140 KCl, 1 MgCl₂, 3 Mg-ATP, 10 HEPES, 5 EGTA, 5 NaCl, and 300 nM free Ca²⁺ calculated using the free WebMax software (www.stanford.edu/~cpatton/webmaxcS), and was adjusted to pH 7.25 with KOH. Ca²⁺-clamped CsCl pipette solution contained (in mM): 140 CsCl, 1 MgCl₂, 3 Mg-ATP, 10 HEPES, 5 EGTA, 5 NaCl, and 600 nM free Ca²⁺calculated using the free WebMax software and was adjusted to pH 7.25 with CsOH. Normal Tyrode's (NT) bath solution contained (in mM): 145 NaCl, 3.6 KCl, 1 MgCl₂, 1.3 CaCl₂, 10 HEPES, 5 glucose, 10 sucrose and was adjusted to pH 7.4 with NaOH. NMDG-Cl bath solution contained: 150 NMDG-Cl, 10 HEPES, 10 glucose, 1 MgCl₂, 1.3 CaCl₂ and was adjusted to pH 7.4 with NMDG-OH. Ionomycin, 1-ethyl-2-benzimidazolinone (1-EBIO) and 1-[(2-chlorophenyl) diphenylmethyl]-1H-pyrazole (TRAM-34) were purchased from Sigma-Aldrich. T16Ainh-A01, a specific inhibitor of Ano-1, was purchased from TOCRIS Biosciences. The chemicals were initially dissolved in dimethyl sulfoxide (DMSO), and were diluted to the bath solution for experiments. The amount of DMSO in the bath solution was kept below 0.1%.

Yin M.Z., Park S.W., Kang T.W., Kim K.S., Yoo H.Y., Lee J., Hah J.H., Sung M.H, & Kim S.J. (2015). Activation of K+ channel by 1-EBIO rescues the head and neck squamous cell carcinoma cells from Ca2+ ionophore-induced cell death. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 20(1), 25-33.

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Pharmacological Modulation of Organ Baths

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The following drugs were dissolved as stock solutions in water or ethanol before being diluted to the final concentrations in organ baths: tetrodotoxin (TTX, 0.6 M; Alamone Laboratories; Jerusalem, Israel), hexamethonium (100 M), hyoscine (1 M), N -nitro-L-arginine (L-NNA, 100 M), nicardipine (3 M), nifedipine (2 M), and 1-[(2-chlorophenyl)diphenylmethyl]-1H-pyrazole (TRAM-34, 3 M) all from Sigma-Aldrich, Castle Hill, Australia, and 5,6-dichloro-1ethyl-1,3-dihydro-2H-benzimidazol-2-one (DCEBIO, 3 M; Tocris; In Vitro Technologies, Noble Park, Australia).

Costa M., Keightley L.J., Wiklendt L., Hibberd T.J., Arkwright J.W., Omari T., Wattchow D.A., Brookes S.J.H., Dinning P.G, & Spencer N.J. (2019). Identification of multiple distinct neurogenic motor patterns that can occur simultaneously in the guinea pig distal colon. American journal of physiology. Gastrointestinal and liver physiology, 316(1).

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