Permeability assays were conducted as previously described (26 (link)). HUVECs were plated onto gelatin-coated 3.0-μm polycarbonate Transwell membranes (Corning Costar) and cultured for 4 days with medium changes every day. Cells were serum-starved for 2 hours in phenol red–free Dulbecco’s modified Eagle’s medium/F12 and treated as described for experiments. HRP was added to the upper chambers at a final concentration of 1.5 μg/ml, and after 30 min, the filters were removed and medium was harvested from the lower chamber. HRP leak was determined by absorbance at 450 nm after incubation with 0.5 mM guaiacol, 50 mM Na2HPO4, and 0.6 mM H2O2 for 15 to 30 min.
Gelatin coated 3.0 μm polycarbonate transwell membranes
Manufactured by Corning
Gelatin-coated 3.0-μm polycarbonate Transwell membranes are a type of lab equipment designed for cell culture applications. They feature a polycarbonate membrane with a pore size of 3.0 micrometers, coated with gelatin to promote cell attachment and growth.
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Lab products found in correlation
2 protocols using gelatin coated 3.0 μm polycarbonate transwell membranes
1
Endothelial Barrier Permeability Assay
2
Quantifying Endothelial Permeability Assay
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Permeability assays were conducted as previously described (26 (link)). HUVECs were plated onto gelatin-coated 3.0-μm polycarbonate Transwell membranes (Corning Costar) and cultured for 4 days with medium changes every day. Cells were serum-starved for 2 hours in phenol red–free Dulbecco’s modified Eagle’s medium/F12 and treated as described for experiments. HRP was added to the upper chambers at a final concentration of 1.5 μg/ml, and after 30 min, the filters were removed and medium was harvested from the lower chamber. HRP leak was determined by absorbance at 450 nm after incubation with 0.5 mM guaiacol, 50 mM Na2HPO4, and 0.6 mM H2O2 for 15 to 30 min.
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