Migration was measured using Matrigel-free transwell plates (Corning, Midland, Michigan, USA) with an 8 μm porous membrane, and invasion was measured using transwell plates coated with Matrigel. Cells as described above were separately seeded into the upper chambers of the transwell at a density of 1.0 × 105 cells per well. The subsequent operation follows the previous description [29 (link)].
Matrigel free transwell plates
Manufactured by Corning
Sourced in United States
Matrigel-free Transwell plates are a type of cell culture insert designed for studying cell migration, invasion, and permeability. They feature a porous membrane that allows cells to migrate from the upper chamber to the lower chamber. The plates do not contain Matrigel, a commonly used extracellular matrix, allowing for a more controlled environment for these types of experiments.
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3 protocols using matrigel free transwell plates
1
Transwell Assay for Cell Migration and Invasion
2
Transwell Assay for Cell Migration and Invasion
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Migration was measured using Matrigel-free Transwell plates (Corning, Inc.) containing an 8-µm porous membrane, while invasion was measured using Transwell plates precoated with 25% Matrigel at 37°C for 30 min. In total, 1×105 A-498 cells stably expressing shRGS20 or shctrl were plated in the upper chambers of the Transwell plates in 100 µl DMEM without FBS. A total of 500 µl DMEM with 10% FBS was plated in the lower chambers of the Transwell plates. After 24 h of incubation 37°C, migrating or invading cells were stained with 0.5% crystal violet at room temperature for 30 min, and then the cells in six random fields were photographed and counted using a light microscope (Motic Incorporation, Ltd.; magnification, ×100) and ImageJ v1.48 (National Institutes of Health), respectively.
3
Transwell Migration and Invasion Assay
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Migration was measured using Matrigel-free Transwell plates (Corning, Midland, MI, USA) with an 8 μm porous membrane. For invasion assay, membrane of the upper chambers was coated with Matrigel (BD Biosciences, Sparks, MD, USA) before use. In total, 1 × 105 cells were plated in the upper chambers of the Transwells. After a 24-h incubation, migrating or invading cells were stained with 0.5% crystal violet, then photographed at 200× and counted in five random fields. CXCR7 inhibitor CCX771 was a courtesy from ChemoCentryx (Mountain View, CA, USA).
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