For the motility assays, 5 × 104 cells were suspended in the migration medium (medium without FBS), and were plated in the top chamber without the Matrigel-coated membrane (24-well insert; 8-μm pore size; BD Biosciences). The lower compartment was filled with 600 μl of a medium containing 30% FBS as a chemo-attractant. Following incubation for 16 h at 37 °C and 5% CO2 in a humidified incubator, the cells on the lower surface of the filter were fixed in 4% formaldehyde for 20 min, then stained with Giemsa stain for 10 min at room temperature, and examined by light microscopy.
Matrigel coated membrane 24 well insert 8 μm pore size
Manufactured by BD
Sourced in United States
Matrigel-coated membrane (24-well insert; 8-μm pore size) is a laboratory equipment product. It consists of a membrane with a pore size of 8 micrometers, which is coated with Matrigel, a basement membrane extract derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells.
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Lab products found in correlation
2 protocols using matrigel coated membrane 24 well insert 8 μm pore size
1
Cell Motility Assay Protocol
2
Transwell Chamber Assay for Cell Migration
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To examine the migratory ability of cells in vitro, transwell chamber assay was used. A total of 2×105 HCT116 cells were placed in the upper compartment of a 24-well transwell unit with 8 μm polycarbonate nucleopore filters (Corning Costar, Cambridge, MA, USA). For the invasion assay, 2×105 cells were plated in the top chamber with a Matrigel-coated membrane (24-well insert, 8 μm pore size; BD Biosciences, Franklin Lakes, NJ, USA). Cells were plated in the medium without serum, and the medium supplemented with 10% serum was used as a chemoattractant in the lower chamber. The cells were incubated for 24–36 h. Cells that did not invade through the pores, were removed by a cotton swab. Filters were fixed with 90% ethanol, stained with 0.1% crystal violet (Beyotime Institute of Biotechnology, Shanghai, China), and photographed. Cell numbers were counted.
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