Krebs buffer contained (in mM) 146.9 NaCl, 4.7 KCl, 2 CaCl2·2H2O, 1.2 MgSO4, 1.2 NaH2PO4·H2O, 3 NaHCO3, 1.5 NaHEPES, and 5 D-glucose (pH = 7.4). Krebs-BSA buffer was prepared with the addition of 0.5 % bovine serum albumin. During cannulation, Krebs-BSA buffer was present both luminally and abluminally, but during the experiment, the bath solution was constantly exchanged with Krebs solution without albumin. All chemicals and drugs were purchased from Sigma-Aldrich (St. Louis, MO), with exception of BSA (United States Biochemicals; Cleveland, OH), MgSO4, Na-HEPES (ThermoFisher Scientific; Pittsburgh, PA), iberiotoxin, ODQ, NS11021 (Tocris Bioscience, Bristol, UK) and apamin (Alomone Labs, Jerusalem, Israel). Sodium NONOate, acetylcholine, Rp-8-Br-PET-cGMPS, iberiotoxin and apamin were dissolved in distilled water. Glibenclamide, ODQ and NS11021 were dissolved in DMSO and the total amount of DMSO was set below 0.4 %, which was determined in separate protocols to be the threshold vasoactive dose of DMSO. Penitrem A was dissolved in methanol, which by itself had no effect on contraction at the concentrations used.
Ns11021
Manufactured by Bio-Techne
Sourced in United States
NS11021 is an analytical tool manufactured by Bio-Techne. It is designed to analyze and quantify the presence and concentration of specific analytes in biological samples.
Automatically generated - may contain errors
4 protocols using ns11021
1
Krebs Buffer Vasodilation Protocol
2
Investigating BK Channel Modulation
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All chemicals were bath applied through a peristaltic pump system at a steady perfusion and suction rate. The time required for the solution to flow to the recording slice through stopcocks was approximately 1 minute. All drugs, including paxilline (a specific BK channel blocker28 , 29 ), were purchased from Sigma‐Aldrich (St. Louis, MO, USA), except NS11021 (a specific BK channel opener27 , 30 , 31 ) and TTX (a sodium channel blocker), which were bought from Tocris Bioscience (Bristol, UK). All chemicals were dissolved in dimethyl sulfoxide (except TTX that was dissolved in distilled water) at 1000 times final concentration, stored at −20°C, and diluted to the final concentration in an oxygenated ASCF solution immediately before use.
3
In Vitro and In Vivo Evaluation of BMS-191011 and NS11021 in TNBC
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BMS-191011 (Tocris) and NS11021 (Tocris) were prepared in 10 mM DMSO stock. The working concentrations of 10-50 μM contained 1-5 μl of DMSO in 1 mL DMEM medium, resulting in 0.1–0.5% of DMSO, which did not affect TNBC cells (Fig. S7 ). For testing in vivo effects of MDA-MD-231, 3 μl of 10 mM (equivalent to 186.82 ng) stock was added to 1 mL PBS, 50 μl was administrated directly into the xenograft grown in mouse via during day time in the animal imaging facility. For testing adverse effects of the drug, tail-vein injection was used.
4
In Vivo Evaluation of BMS-191011 and NS11021 in TNBC
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BMS-191011 (Tocris) and NS11021 (Tocris) were prepared in 10mM DMSO stock. The working concentrations of 10-50mM contained 1-5ml of DMSO in 1 mL DMEM medium, resulting in 0.1-0.5% of DMSO, which did not affect TNBC cells (Figure S7). For testing in vivo effects of MDA-MD-231, 3ml of 10mM (equivalent to 186.82ng) stock was added to 1mL PBS, 50ml was administrated directly into the xenograft grown in mouse via during day time in the animal imaging facility. For testing adverse effects of the drug, tail-vein injection was used.
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