HUVECs were cultured in Transwell chambers (0.4 μm pore polycarbonate filters; Costar, Cambridge, Miss.). After reaching confluence, the medium was replaced with the previously-collected conditioned medium (0.3 ml in the upper chamber and 1 ml in the lower chamber). Horseradish peroxidase molecules (type VI-A, 44 kDa; Sigma-Aldrich) at a concentration of 0.126 μM were then added to the upper compartment. After incubating for 1 hr, the medium in the lower compartment was assayed for enzymatic activity using a photometric guaiacol substrate assay (Sigma-Aldrich).
Transwell chambers 0.4 μm pore polycarbonate filters
Manufactured by Corning
Sourced in United States
Transwell chambers are laboratory equipment used for cell culture experiments. They consist of a polycarbonate membrane with 0.4 μm pores, which separates the upper and lower chambers. This configuration allows for the study of cell migration, permeability, and other cellular interactions across a semi-permeable barrier.
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Lab products found in correlation
2 protocols using transwell chambers 0.4 μm pore polycarbonate filters
1
Permeability Assay for HUVECs
2
HRP Permeability Assay in HUVECs
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HUVECs were cultured in Transwell chambers (0.4 μm pore polycarbonate filters; Costar, Cambridge, MA, USA). After reaching confluence, the medium was replaced with the CM (0.3 mL in the upper chamber and 1 mL in the lower chamber). Horseradish peroxidase molecule (Sigma-Aldrich, Saint Louis, MO, USA) was added to the upper compartment at a concentration of 0.2 µM. After incubation for 1 h, the medium in the lower compartment was assayed for enzymatic activity while using a photometric guaiacol substrate assay (Sigma-Aldrich).
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