Ucl1684

Manufactured by Bio-Techne

Sourced in United Kingdom

UCL1684 is a laboratory equipment product offered by Bio-Techne. It serves as a core function in various research and analysis applications.

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Lab products found in correlation

10 protocols using ucl1684

Isolation and Characterization of Porcine Arterial Endothelial Cells

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Porcine arterial endothelial cells were isolated as described previously [48 (link)]. In brief, porcine coronary arteries were cut open longitudinally and incubated for 30 min in phosphate-buffered saline w/o Ca²⁺/Mg²⁺ containing trypsin/EDTA (0.25%/0.02%, w/v). Thereafter, the luminal surface was gently scrapped with the tip of a 100 μL pipette tip. Detached endothelial cells were aspirated and plated on cover slips in Modified Eagle Medium supplemented with 10% newborn calf serum and penicillin/streptomycin (all from Biochrom KG, Berlin, Germany). Whole cell currents were recorded with an EPC10-USB patch-clamp amplifier and the PatchmasterTM software (HEKA Electronics, Germany) using voltage-ramps (−100 to 100 mV, 1 sec, filter 1000 Hz). K⁺ outward currents were quantified at a potential of 0 mV before (w/o), after addition of CyPPA, and after addition of UCL1684 (both from Tocris Bioscience). The KCl-pipette solution was composed of (in mM): 140 KCl, 1 MgCl₂, 2 EGTA, 1.71 CaCl₂ (1 μM [Ca²⁺]_free), and 5 HEPES (adjusted to pH 7.2 with KOH). The bath solutions was composed of (in mM): 140 NaCl, 5 KCl, 1 MgSO₄, 1 CaCl₂, 10 glucose and 10 HEPES (adjusted to pH 7.4 with NaOH).

Simonsen U., Winther A.K., Oliván-Viguera A., Comerma-Steffensen S., Köhler R, & Bek T. (2019). Extracellular l-arginine Enhances Relaxations Induced by Opening of Calcium-Activated SKCa Channels in Porcine Retinal Arteriole. International Journal of Molecular Sciences, 20(8), 2032.

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Pharmacological Modulation of KCa Channels

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Pharmacological modulators of K_Ca2.x and K_Ca3.x channels were purchased from Tocris Bioscience; CyPPA (Cat. no. 2953), UCL1684 (Cat. no. 1310) and NS6180 (Cat. no. 4864). NS8593 (Cat. no. N2538) was purchased from Sigma‐Aldrich, UK.

Abdulkareem Z.A., Gee J.M., Cox C.D, & Wann K.T. (2015). Knockdown of the small conductance Ca2+‐activated K+ channels is potently cytotoxic in breast cancer cell lines. British Journal of Pharmacology, 173(1), 177-190.

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Pharmacological Modulation of Gamma Oscillations

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Physostigmine (Phys), NS309, NS6180, UCL1684, NS19504, Penitrem A, XE991, ICA110381, SAK3, and NNC55-0396, CNQX, D-APV, and TTX were obtained from Tocris Bioscience (Bristol, UK). Acetylcholine (ACh) was purchased from Sigma-Aldrich (Taufkirchen, Germany), TTA-P2 from Alomone Labs (Jerusalem, Israel), picrotoxin from Abcam (Cambridge, UK). Drugs were dissolved in water or DMSO and further diluted with ACSF to achieve the respective drug concentrations. Final DMSO concentration was held below 0.02% or DMSO control experiments were carried out. We did not investigate the effect of DMSO on gamma oscillation power systematically; however, it is known that DMSO reduces neuronal excitability (Tamagnini et al., 2014 (link)), which can explain the lower normalized DMSO control values. In patch clamp experiments, the final DMSO concentration was 0.2%.

Klemz A., Wildner F., Tütüncü E, & Gerevich Z. (2022). Regulation of Hippocampal Gamma Oscillations by Modulation of Intrinsic Neuronal Excitability. Frontiers in Neural Circuits, 15, 778022.

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Ion Channel Modulators in Cellular Assays

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Calcium ionophore A23187 (1 μM), 4-α-phorbol 12,13-didecanoate (4αPDD) (1 μM), GSK1016790A (100 nM), RN1734 (5 μM), HC067047 (300 nM), UCL-1684 (30 nM), TRAM-34 (30 nM) and iberiotoxin (IbTX) (1, 10, 30 and 100 nM) were all dissolved in DMSO and diluted to a final working concentration of no more than 0.01% DMSO (0.01% DMSO had no effect alone). Calcium ionophore A23187, TRPV4 channel agonist 4αPDD (Vincent et al., 2009 (link)), TRPV4 channel agonist GSK1016790A (Thorneloe et al., 2008 (link)) and BK channel inhibitor IbTX (Candia et al., 1992 (link)) were sourced from Sigma-Aldrich, and TRPV4 channel antagonist RN1734 (Vincent et al., 2009 (link)), TRPV4 channel antagonist HC067047 (Everaerts et al., 2010 (link)), SK channel inhibitor UCL-1684 and IK channel inhibitor TRAM-34 (Nehme et al., 2012 (link)) were sourced from Tocris, Bristol, UK.

Feetham C.H., Nunn N., Lewis R., Dart C, & Barrett-Jolley R. (2015). TRPV4 and KCa ion channels functionally couple as osmosensors in the paraventricular nucleus. British Journal of Pharmacology, 172(7), 1753-1768.

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Assay for Chemical Compound Procurement

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All chemicals were purchased from Sigma-Aldrich (Sigma Aldrich Chimie S.A.R.L., St.-Quentin-Fallavier, France) except TRAM-34, UCL-1684, NS-398, and SC-560, which were bought from Tocris (Bio-Techne, Abingdon, UK).

Gaertner S., Auger C., Farooq M.A., Pollet B., Khemais-Benkhiat S., Niazi Z.R., Schrevens S., Park S.H., Toti F., Stephan D, & Schini-Kerth V.B. (2020). Oral Intake of EPA:DHA 6:1 by Middle-Aged Rats for One Week Improves Age-Related Endothelial Dysfunction in Both the Femoral Artery and Vein: Role of Cyclooxygenases. International Journal of Molecular Sciences, 21(3), 920.

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Ion Channel Modulator Preparation

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10 mM NS309 (Cayman Chemical, Ann Harbor, MI, USA), 5 mM UCL1684 (Tocris/Bio-Techne, Minneapolis, MN, USA), and 5 mM HMR1556 (Sigma-Aldrich, St. Louis, MO, USA) stock solutions were prepared in DMSO. 1 mM stock solution of apamin (Alomone Labs, Jerusalem, Israel)) and 1 mM stock of E4031 hydrochloride (Cayman Chemical, Ann Harbor, MI, USA) was prepared in deionized water. Stock solutions were diluted to final concentration in external solutions. Corresponding amount of DMSO was added to control solution (final DMSO concentration ≤0.06%).

Kanaporis G, & Blatter L.A. (2022). Activation of small conductance Ca2+-activated K+ channels suppresses Ca2+ transient and action potential alternans in ventricular myocytes. The Journal of physiology, 601(1), 51-67.

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Coronary Flow Regulation Pharmacology

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5-HT, adenosine, histamine, indomethacin, tetraethylammonium chloride (TEA), L-NAME, and dimethyl sulfoxide (DMSO) were purchased form Sigma-Aldrich (St. Louis, Missouri, USA). SB269970, R96544, 5-carboxamindotryptamine (5-CT), TRAM-34, and UCL1684 were purchased from Tocris Bioscience (Bristol, United Kingdom). Quinacrine, penitrem A, paxilline, and SKF525A were purchased from Cayman Chemical Co. (Ann Arbor, Michigan, USA).
The perfusion solution used in the present study was modified Tyrode’s solution (in mM): 119.7 NaCl, 23.8 NaHCO₃, 5.6 Glucose, 1.2 CaCl₂, 1.1 MgCl₂, 0.3 NaH₂PO_4, and 5.0 KCl. In the experiments testing the effects of the inhibitors of arachidonic acid metabolism and the blockers of Ca²⁺-activated K⁺ channels on 5-HT-induced coronary flow increases, L-NAME 10 μM was added to the perfusion solution and existed throughout the experiments.

Chang Chien C.C, & Su M.J. (2015). 5-hydroxytryptamine has an endothelium-derived hyperpolarizing factor-like effect on coronary flow in isolated rat hearts. Journal of Biomedical Science, 22(1), 42.

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Pharmacological Modulation of hMC Electrophysiology

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During this study, we used a glutamatergic receptor antagonist (10 µM cyanquixaline, or CNQX, Millipore-Sigma) and a GABAA chloride channel blocker (10 mM picrotoxin, or PTX, Abcam) to abolish synaptic activity during our recordings. Furthermore, we also used a Ca 2+activated K + (SK) channel blocker, 30 µM UCL1684 (Tocris) to study the after-hyperpolarizing potential (AHP) in hMCs.

Trinh A., Girardi‐Schappo M., Béïque J., Longtin A., & Maler L. (2022). Dentate gyrus mossy cells exhibit sparse coding via adaptive spike threshold dynamics.

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Concentration-Dependent Inhibition Quantification

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Apamin and UCL1684 (both Bio-Techne) were diluted to 100 μM in ddH₂O and DMSO, respectively, with aliquots stored at −20 °C. Aliquots were thawed on the day of use and diluted further in the external solution described above. Aliquots were sequentially added to the superfusate to obtain concentrations ranging from 10 pM to 300 nM. Concentration–inhibition relationships were fit and a standard Hill equation in the form of:

\frac{I}{I_{c o n t}} = \frac{A_{\max}}{1 + 10^{({L o g I C}_{50} - x) n_{h}}}

where I_cont and I are the current magnitude in the absence and presence of a given concentration of drug (

x

; in Log(M)) respectively.

A_{\max}

is the maximal inhibition, IC₅₀ is the concentration of an inhibitor which inhibits 50% of the sensitive current, and

n_{h}

is the Hill coefficient (using Prism 9.2.0 (GraphPad). Where data were best fit by the sum of two Hill equations, modified equations were totaled in which

A_{\max}

was multiplied by the fraction of the inhibition (

A_{f r a c}

) which each component (

a

and

b

) accounted for:

\frac{I}{I_{c o n t}} = \frac{A_{\max} \times A_{f r a c, a}}{1 + 10^{({L o g I C}_{50, a} - x) n_{h, a}}} + \frac{A_{\max} \times A_{f r a c, b}}{1 + 10^{({L o g I C}_{50, b} - x) n_{h, b}}}

Butler A.S., Hancox J.C, & Marrion N.V. (2022). Preferential formation of human heteromeric SK2:SK3 channels limits homomeric SK channel assembly and function. The Journal of Biological Chemistry, 299(1), 102783.

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Ion Channel Modulators Protocol

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Aliquots of stock solutions of all drugs were stored at -20°C and diluted 1:1000 in external solution on the day of use. ACh (Sigma-Aldrich) and apamin (Bio-techne) were dissolved in H₂O. UCL1684 (Bio-techne) and E-4031 (Caymen chemicals) were dissolved in DMSO. 100 nM apamin, 100 nM UCL1684 and 1 µM E-4031 was used to maximally inhibit I_SK and I_Kr respectively^{95 (link),96 (link)}. 1 µM ACh was used to fully activate I_K,ACh^{97 (link)}.

Butler A.S., Ascione R., Marrion N.V., Harmer S.C, & Hancox J.C. (2024). In situ monolayer patch clamp of acutely stimulated human iPSC-derived cardiomyocytes promotes consistent electrophysiological responses to SK channel inhibition. Scientific Reports, 14, 3185.

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