Matrigel-coated nucleopore filter inserts in a 24-well transwell chamber (Corning Biocoat, New York, USA) were used for the invasion assays. Cells (treated or untreated with VS-6063) were seeded at a density of 40,000 cells/well into the upper part of the Matrigel-coated filter, and RPMI-1640 supplemented with 10% heat-inactivated fetal bovine serum and rhCXCL-12 (R&D Systems, Wiesbaden, Germany) (1 ng/mL) was added to the lower part. After 24 h, the cells that had migrated through the Matrigel and the 8-μm pore-size membrane were fixed, stained, and counted under a light microscope.
Matrigel coated nucleopore filter inserts in a 24 well transwell chamber
Manufactured by Corning
Sourced in United States
Matrigel-coated nucleopore filter inserts in a 24-well transwell chamber. This product provides a platform for cell migration and invasion assays.
Automatically generated - may contain errors
2 protocols using matrigel coated nucleopore filter inserts in a 24 well transwell chamber
1
Transwell Invasion Assay with Matrigel
2
Quantitating CLL Cell Invasion
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Matrigel-coated nucleopore filter inserts in a 24-well transwell chamber (Corning) were used for the invasion assays. Cells were seeded at a density of 5 × 105 cells per well in Medium 199 supplemented with 10% FCS and IL-4 (5 ng/mL). PBMCs from six patients were pre-treated for 2 h with a range of defactinib concentrations (0.5 µM, 1 µM, 5 µM) or with an equivalent amount of DMSO. The cells were then added to the apical chamber of the invasion assay plate for 24 h (in defactinib + media) and encouraged to invade towards a CXCL12 gradient created by adding 700 µL complete media, supplemented with 10 ng/mL CXCL12 (R&D Systems, Minneapolis, MN, USA), into the basolateral chamber. Invading CD19+/CD5+ cells were quantitated by volumetric counting using the Cytoflex LX flow cytometer (Beckman Coulter).
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