Transepithelial electrical resistance (TEER) is a widely accepted quantitative technique to measure the barrier integrity [17 (link)]. A higher TEER indicates a better barrier function. The in vitro TEER measurement was performed following the protocol described previously. Briefly, 8 keratinocytes were seeded on the polyester filter membrane of a transwell device (24-well format, 0.4 mm pore size, Corning, USA) and cultured in the diluted BG extracts and the control medium, respectively. The TEER value was measured using a Millicell ERS-2 epithelial volt-ohmmeter (Millipore, USA) on days 1, 3, 7. The net value of each group was calculated by subtracting the background value, which was measured on the cell-free filter membrane, from the value of each group [18 ].
24 well format
Manufactured by Corning
Sourced in United States
The 24-well format is a laboratory equipment commonly used in cell culture and assay applications. It provides a standardized multi-well layout that facilitates parallel experiments and sample processing. The product offers a convenient and efficient platform for various biological and biochemical analyses.
Automatically generated - may contain errors
Lab products found in correlation
Transwell device, by Corning (1 mentions)
Millicell ers 2 epithelial volt ohm meter, by Merck Group (1 mentions)
Evans blue dye, by Merck Group (1 mentions)
Fitc dextran, by Thermo Fisher Scientific (1 mentions)
Formamide, by Merck Group (1 mentions)
Crystal violet, by Merck Group (1 mentions)
8.0 m pore polycarbonate membrane insert, by Corning (1 mentions)
4 protocols using 24 well format
1
Quantifying Epithelial Barrier Function via TEER
2
Quantifying Endothelial Permeability in Rats
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Rats were injected 1% evans blue (EB) dye (Sigma) (6 mL/kg) in PBS/4% bovine serum albumin (BSA) via tail vein. One hour later, animals were sacrificed and the lungs were dissected after completely flushing out residual intravascular dye with PBS. Dissected lungs were photographed and homogenized in formamide (Sigma), then incubated at 37°C for 24 hours. After centrifuge, the supernatant was collected and measured at 620 nm on plate spectrophotometer.
To assess endothelial monolayer permeability, transwells (0.4 µm; 24‐well format; Corning) were plated with rPAECs at the density of 5 × 103/well. After transfecting rPAECs with siRNA or plasmids and subsequent MCTP or vehicle treatments, FITC‐dextran (Invitrogen) was added to the upper chamber and incubated at 37°C for 1 hour. Medium (20 µL) was collected from the lower chamber and measured with excitation at 488 nm and emission at 520 nm on plate spectrophotometer.
To assess endothelial monolayer permeability, transwells (0.4 µm; 24‐well format; Corning) were plated with rPAECs at the density of 5 × 103/well. After transfecting rPAECs with siRNA or plasmids and subsequent MCTP or vehicle treatments, FITC‐dextran (Invitrogen) was added to the upper chamber and incubated at 37°C for 1 hour. Medium (20 µL) was collected from the lower chamber and measured with excitation at 488 nm and emission at 520 nm on plate spectrophotometer.
3
Transwell Cell Migration Assay
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3×104 transfected GC cells were cultured in 400 μl serum-free media and seeded in the upper part of each transwell chamber in a 24-well format (Corning, USA). 800 μl of normal MEM medium containing 10% FBS was added as a chemoattractant in the bottom chamber. Similarly, approximately 6×104 cells suspended in 400μl serum-free media were seeded independently in the top chamber coated with matrigel for invasion assays. After incubation for 24-48 h, the cells on the upper part of the chamber were removed with a cotton applicator, whereas the cells migrated through the membrane were stained with crystal violet. The migrated cells on the bottom surface of the membrane were photographed and counted on an inverted microscope. The migrated cells were counted in five random fields under a light microscope (200×magnification) and the average number of five fields was calculated. All assays were performed in triplicate and repeated three times.
4
Transwell Cell Migration Assay
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Assay was performed in 6.5 mm transwell with 8.0 µm pore polycarbonate membrane insert in a 24-well format (Corning Inc.; Corning, NY, USA). HCT116 cells were seeded into upper chambers at 1 × 105 cells in serum free-medium and incubated with indicated condition for 24 h for cell migration. HUVEC cells were pretreated with indicated condition for 24 h and then seeded into upper chambers at 1 × 105 cells in EBM2 basal medium and incubated with indicated condition for 6 h for migration. Membranes then fixed with 10% neutral buffered formalin at room temperature for 10 min and stained with crystal violet (Sigma-Aldrich; St. Louis, MO) at room temperature for 20 min. Membranes were washed with water twice and no-migrated cells were scrap off by cotton stick. crystal violet of migrated cells was dissolved using 33% acetic acid and then detected at wavelength 600 nm.
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