A253 cells were cultured to confluence in the inner chambers of 12 mm transwell inserts with 0.4 μM pore-size filters (Corning Life Sciences). The TEER was measured using an EVOM voltameter with an ENDOHM-12 (World Precision Instruments, Sarasota, FL, USA) on a heating plate (Fine, Tokyo, Japan) adjusted to 37 °C. The values were expressed in standard units of ohms per square centimeter and presented as the mean ± S.D. For calculation, the resistance of blank filters was subtracted from that of filters covered with cells.
Endohm 12
Manufactured by World Precision Instruments
Sourced in United States, Japan
The ENDOHM-12 is a laboratory instrument designed to measure the electrical properties of materials. It is capable of measuring the impedance, resistance, and capacitance of samples within a specific frequency range.
Automatically generated - may contain errors
Lab products found in correlation
Evom voltmeter, by World Precision Instruments (5 mentions)
Evom voltameter, by World Precision Instruments (3 mentions)
0.4 μm pore size filters, by Corning (2 mentions)
12 mm transwell inserts with 0.4 m pore size filters, by Corning (2 mentions)
Evom2, by World Precision Instruments (2 mentions)
Transwell insert, by Corning (2 mentions)
3 μm pore size filters, by Corning (2 mentions)
Transwell 0.4 μm pore size filters, by Corning (1 mentions)
0.4 μm pore filters, by Corning (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
13 protocols using endohm 12
1
Transwell Barrier Integrity Evaluation
2
Transepithelial Voltage and Resistance Measurements
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Transepithelial voltage and resistance measurements were performed using Endohm chambers (ENDOHM‐24SNAP or ENDOHM‐12) from World Precision Instruments (WPI) connected to an EVOM2 epithelial voltohmmeter from WPI in voltage‐clamp short‐circuit mode. Short circuit equivalent current (Ieq) was calculated from voltage and resistance read outs using Ohm's law, I = V/R. We report values of Ieq relative to the control group. More than 95% of Ieq was inhibited by 10 μmol/L apical amiloride and thus is considered to be proportional to the activity of ENaC in the apical plasma membrane. Before collecting electrophysiological read outs, 100 μL of apical differentiation media was gently added to cell culture inserts in addition to the 100 μL of apical differentiation media already present to ensure electrical contact between the upper electrodes and apical fluid.
3
Transwell TEER Measurement in hTERT-HNECs
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hTERT-transfected HNECs were cultured to confluence in the inner chambers of 12-mm Transwell inserts with 0.4-µm pore-size filters (Corning Life Sciences; Tewksbury, MA, USA). TEER was measured using an EVOM voltameter with an ENDOHM-12 (World Precision Instruments; Sarasota, FL, USA) on a heating plate (Fine; Tokyo, Japan) adjusted to 37 °C. The values were expressed in standard units of ohms per square centimeter and presented as the mean ± S.D. For calculation, the resistance of blank filters was subtracted from that of filters covered with cells.
4
Primary Pulmonary Endothelial Cell Culture
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Primary human pulmonary microvascular endothelial cells (PromoCell C-12281) were grown on transwells (Costar, 3460; for transendothelial electrical resistance measurements, TEER) or glass coverslips (for immunofluorescent imaging). TEER was measured using Endohm-12 from World Precision Instruments. For imaging, cells were fixed with 4% PFA and counterstained with DAPI. Confocal imaging was performed with a Quorum TIRF/SD microscope.
5
TEER Measurement of Cell Monolayers
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TEER measurements were performed according to a protocol adapted from a publication by the L’Oreal Research group89 . TEER was measured using the EVOM2 and the Endohm-12 (World Precision Instruments) and D-PBS (Ca2+, Mg2+) as an electrolyte solution. The day of TEER measurement, 0.3 ml of D-PBS was added onto the apical side of the insert and left to equilibrate for 15 min at 37 °C, and 2.5 ml of D-PBS were added inside an EndOhm Chamber. Following TEER measurements, D-PBS was removed from the apical side to restore the air-liquid interface.
6
Transepithelial Electrical Resistance Measurement
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The cells were cultured to confluence in the inner chambers of 12‐mm Transwell 0.4‐μm pore‐size filters (Corning Life Science). Transepithelial electrical resistance (TEER) was measured using an EVOM voltmeter with an ENDOHM‐12 (World Precision Instruments, Sarasota, FL) on a heating plate (Fine, Tokyo, Japan) and was monitored using a cellZscope (nanoAnalytics, Münster Germany), a computer controlled automated multiwell device (12 wells). The values were expressed in standard units of ohms per square centimeter and presented as the mean ± SD of triplicate experiments. For calculation, the resistance of blank filters was subtracted from that of filters covered with cells.
7
Transwell Assay for Transepithelial Resistance
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Cells were cultured to confluence on the inner chambers of 12-mm Transwell inserts with 0.4-μm pore filters (Corning Life Sciences, NY, USA). TER was measured using an EVOM voltmeter with an ENDOHM-12 (World Precision Instruments, Sarasota, FL, USA). Data are expressed in Ω/cm2 and presented as the mean ± S.D. of triplicate experiments. For calculation, the resistance of blank filters was subtracted from that of filters covered with cells.
8
Assessing Epithelial Barrier Integrity
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Trans-epithelial electrical resistance: This method evaluates barrier integrity of epithelial cells grown in monolayer; by describing the impedance of barrier-forming cell cultures. Briefly, electrodes are placed on both sides of the cellular barrier and an electric current is applied. The resulting current established in the circuit is measured and trans-epithelial electrical resistance (TEER) is calculated. The higher the TEER, the better the membrane integrity[32 ]. In this study, HT-29, HT-29 Cx43D, and HT-29 Cx43- cells were cultured on Transwell® inserts with 0.4 μm-pore size filters (Corning, Corning, NY, United States). TEER was measured on confluent cells in the presence or absence of 2% dextran sulfate sodium (DSS) using an EVOM voltmeter with an ENDOHM-12 (World Precision Instruments, Sarasota, FL, United States). Electrical resistance was expressed as Ω × cm2. DSS was applied onto the cells to reproduce in vitro the membrane breach it is known to induce in vivo and characterize the loss of membrane integrity. TEER was calculated by subtracting the resistance of blank filters from that of filters covered with a monolayer of parental HT-29, HT-29 Cx43D, or HT-29 Cx43- cells.
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9
Transepithelial Electrical Resistance in Caco-2 Cells
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Caco-2 cells were cultured on Transwell inserts with 3-μm–pore size filters (Falcon® Cell Culture Inserts, Sterile, Corning®). Five days post-confluence, the TEER was measured over a period of 24 h using an EVOM voltmeter with an ENDOHM-12 (World Precision Instruments, Florida USA). Electrical resistance was expressed as Ω × cm2. The TEER was calculated by subtracting the resistance of blank filters from that of filters covered with a monolayer of Caco-2 cells.
10
Transepithelial Electrical Resistance Measurement
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Cells were cultured on Transwell inserts with 3-μm–pore size filters (Corning). Transepithelial electrical resistance was measured using an EVOM voltmeter with an ENDOHM-12 (World Precision Instruments, Sarasota, FL). Electrical resistance was expressed as Ω × cm2. For calculation, the resistance of blank filters was subtracted from that of filters covered with cells.
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