(1,3-dihydro-1-[2-hydroxy-5-(trifluoromethyl)-phenyl]-5-(trifluoromethyl)-2H-benzimidazol-2-one), NS1619 (Tocris), a specific BKCa channel opener; Paxilline (alomone), a BKCa channel blocker; NS1619 and Paxilline were both dissolved in dimethysulfoxide (DMSO), and then diluted in ACSF to the final concentration of 10 μM.
Paxilline
Manufactured by Bio-Techne
Sourced in United Kingdom, United States
Paxilline is a chemical compound that functions as a potassium channel blocker. It is commonly used as a research tool in the study of ion channel regulation and cellular signaling processes.
Automatically generated - may contain errors
Lab products found in correlation
Glibenclamide, by Bio-Techne (4 mentions)
4 aminopyridine 4 ap, by Merck Group (3 mentions)
Phenylephrine pe, by Merck Group (3 mentions)
Acetylcholine, by Bio-Techne (3 mentions)
Indomethacin, by Bio-Techne (2 mentions)
Linopirdine, by Bio-Techne (2 mentions)
Kt5823, by Bio-Techne (2 mentions)
Dpo 1, by Bio-Techne (2 mentions)
Psora 4, by Bio-Techne (2 mentions)
L nna, by Bio-Techne (2 mentions)
Nifedipine, by Merck Group (2 mentions)
Ns1619, by Bio-Techne (1 mentions)
Tetrodotoxin, by Bio-Techne (1 mentions)
Verapamil, by Merck Group (1 mentions)
Glutamate, by Merck Group (1 mentions)
Muscimol, by Merck Group (1 mentions)
Bicuculline methiodide, by Merck Group (1 mentions)
Ns309, by Bio-Techne (1 mentions)
Xe991, by Merck Group (1 mentions)
Horseradish peroxidase conjugated anti rabbit secondary antibody, by Santa Cruz Biotechnology (1 mentions)
19 protocols using paxilline
1
BK Channel Modulation Protocol
2
Reconstitution and Application of Peptides
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Paxilline and tetrodotoxin (Tocris, Bristol, UK) were prepared as 1,000 × stocks in DMSO and water, respectively. β2N, was synthesized as the first 45 amino acids of the β2 subunit (MFIW
3
Pharmacological Modulation of Ion Channels
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Drugs were applied to the perfusion bath or through puff apparatus, as previously described [15 (link)]. Verapamil (20 µM), nickel (100 µM or 200 µM), TEA (2 mM), muscimol (10 µM), bicuculline methiodide (BMI, 10 µM), glutamate (100 µM), NMDA (300 µM), AP5 (50 µM), CNQX (10 µM) were purchased from Sigma, Italy. Paxilline (10 µM) was purchased from Tocris Bioscience UK, and TTX (500 nM) from Alomone Labs, Israel.
4
Biochemical Reagents for Cell Assays
5
Physiological Saline Solution for Mesenteric Artery Myography
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Physiological saline solution (PSS) for surgical isolation of mesenteric arteries and myography experiments containing 6 mM KCl, 112 mM NaCl, 1.18 mM NaHCO3, 1.18 mM MgSO4, 1.18 mM KH2PO4, 1.18 mM CaCl2, and 10 mM glucose was gassed with 21% O2/5% CO2 to pH the solution to approximately 7.4. 60 mM K+-PSS (60K) that was used to test the viability of isolated vessel segments was prepared by equimolar replacement of NaCl with KCl. Dapagliflozin was purchased from Ambeed Inc. (Arlington Heights, IL, USA). Phenylephrine (PE), and 4-aminopyridine (4-AP) and XE 991 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Indomethacin, DPO-1, Linopirdine, Psora-4, Glibenclamide, paxilline, ODQ, KT5823, L-NNA, SNP and acetylcholine (ACh) were purchased from Tocris (Minneapolis, MN, USA). Drug/modulator stocks were prepared by dissolving them in suitable solvents: 4-AP, SNP and PE in distilled water; dapagliflozin, DPO-1, Linopirdine, Psora-4, Indomethacin, Glibenclamide, paxilline, ODQ, KT5823, L-NNA, and ACh in dimethyl sulfoxide (DMSO, final concentration <0.1%). Anti-SGLT2 antibody was purchased from Abcam (Cambridge, UK), and anti-rabbit horseradish peroxidase-conjugated secondary antibody from Santa Cruz Biotechnology (Dallas, TX, USA).
6
Immunofluorescence Staining Protocol
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All primary antibodies used in the present study are listed in Table S7. All secondary antibodies were isotype-specific and conjugated to Alexa Fluor 488- or Alexa Fluor 555 (Thermo Fisher Scientific). dl -2-Amino-5-phosphonopentanoic acid sodium salt (AP-5, #105), Forskolin (#1099), and Paxilline (#2006) were from Tocris. Nifedipine (#N7634), Picrotoxin (#P1675) and Rolipram (#R6520) were from Sigma-Aldrich. DNA oligonucleotide primers used in this study are listed in Table S6. If not specifically stated, all other reagents were of standard quality and from the usual vendors.
7
Experimental Protocols for Potassium Channel Modulators
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TRAM-34 and SKA-31 were kind gifts from Dr. Heike Wulff, Department of Pharmacology, University of California Davis, California, USA. Paxilline was purchased from Tocris Bioscience (Bristol, United Kingdom). TRIZOL reagent was purchased from Invitrogen (United Kingdom). The RNase-Free DNase Set (Qiagen, Germany) was used for DNase digestion. Complementary DNA (cDNA) was synthesized using iScript cDNA Synthesis Kit (Bio-Rad, CA, USA). The new negative-gating modulator of KCa3.1, RA-2 (1,3-phenylenebis(methylene) bis(3-fluoro-4-hydroxybenzoate), was synthesized in house [61 ]
8
Thalidomide and Paxilline Behavioral Assays
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Thalidomide (Tocris Bioscience, Minneapolis, MN, USA) was dissolved to 25 mM in dimethylsulfoxide (DMSO) and then diluted 1:10 in saline for injection. Paxilline (Tocris Bioscience) was dissolved to 10 mM in DMSO and then diluted 1:2,000 in saline for injection. Thalidomide (30 mg/kg) or vehicle (10% (v/v) DMSO) was injected intraperitoneally of 24 h before open field test, tail suspension test, forced swimming test, elevated plus maze test, or the three-chamber social interaction tests. In the passive avoidance test and novel object recognition tests, the same dose of Thalidomide was injected two times before the tests. Paxilline (3 μg/kg) or vehicle (0.05% (v/v) DMSO) was injected intraperitoneally 3 h before the tests.
9
Perforated Patch-Clamp and Ca2+ Imaging Experiments
10
Selective Potassium Channel Blockade in Myoblasts
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The small molecule paxilline (Tocris, Ellisville, MI, USA) and the peptide iberiotoxin (CS Bio, Menlo Park, CA, USA) were used as a selective blocker of KCa1.1.60 (link) Tetraethylammonium (TEA) chloride (Sigma) was used as a nonselective blocker of potassium channels. The α-subunits of KCa1.1 were transduced into myoblasts using the Bacmam expression system as described;42 (link), 61 BacMam vectors expressing GFP were used as controls. Myoblasts were transduced at a multiplicity of infection of 100.
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