Cell migration or invasion was examined using Matrigel-uncoated or Matrigel-coated transwell inserts (8 μm pore size, Corning, USA). U937 and Raji cells (4 × 104 cells/well) were suspended in 150 μL serum-starved culture medium and then added to the upper chambers, and the lower chambers were filled with 700 μL of DMEM medium containing 10% FBS. Twenty-four hours later, cells that had moved through the transwell membrane and invaded the lower chamber were fixed with 70% methanol and stained with 1% crystal violet. The migrated or invaded cells were imaged using a fluorescence microscope and counted in five randomly selected fields.
Matrigel uncoated
Manufactured by Corning
Sourced in United States
Matrigel-uncoated is a gelatinous protein mixture secreted by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is used as a substrate for the in vitro culture of various cell types, including cells from epithelial, endothelial, and neural origins.
Automatically generated - may contain errors
3 protocols using matrigel uncoated
1
Cell Migration and Invasion Assay
2
Evaluating NRSN2 Effects on Breast Cancer Cell Migration and Invasion
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MDA-MB-231 cells were transfected with pRK5-hNRSN2 or siR-NRSN2. Matrigel-uncoated and -coated migration inserts (8-µm pore size; Corning Inc.) were used for migration and invasion assays, respectively.
For the migration assay, MDA-MB-231 cells (1×104) in DMEM were plated into the upper chamber with the non-coated membrane. For the invasion assay, pRK5-hNRSN2 or siR-NRSN2-transfected cells were prepared at a density of 1×104 cells in 500 µl serum-free DMEM in the upper chamber and the lower chamber contained DMEM with 5% FBS. Cells were seeded in the upper chamber of a BD BioCoat Matrigel Invasion Chamber (BD Biosciences) according to the manufacturer's instructions. Following incubation for 48 h, cells were fixed with 4% paraformaldehyde for 1 h at room temperature and stained with 0.1% crystal violet (Sigma-Aldrich; Merck KGaA) for 10 min at 37°C. The membranes were mounted onto a glass slide with antifade mounting medium (cat. no. P0126; Beyotime Institute of Biotechnology), and the number of migrating and invading tumor cells was counted in at least three randomly selected fields under a light microscope (Olympus Corporation) at ×200 magnification.
For the migration assay, MDA-MB-231 cells (1×104) in DMEM were plated into the upper chamber with the non-coated membrane. For the invasion assay, pRK5-hNRSN2 or siR-NRSN2-transfected cells were prepared at a density of 1×104 cells in 500 µl serum-free DMEM in the upper chamber and the lower chamber contained DMEM with 5% FBS. Cells were seeded in the upper chamber of a BD BioCoat Matrigel Invasion Chamber (BD Biosciences) according to the manufacturer's instructions. Following incubation for 48 h, cells were fixed with 4% paraformaldehyde for 1 h at room temperature and stained with 0.1% crystal violet (Sigma-Aldrich; Merck KGaA) for 10 min at 37°C. The membranes were mounted onto a glass slide with antifade mounting medium (cat. no. P0126; Beyotime Institute of Biotechnology), and the number of migrating and invading tumor cells was counted in at least three randomly selected fields under a light microscope (Olympus Corporation) at ×200 magnification.
3
Transwell Migration and Invasion Assays
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Transwell migration and invasion assays were performed using the Matrigel-uncoated or Matrigel-coated transwell chambers (8 μm; Corning, USA) respectively. U937 and Raji cells (4 x 104 cells/well) were suspended in 200 μL serum-free culture medium and then seed into the upper chamber, and 700 μL of DMEM medium containing 2% FBS was added into the lower chamber. Twenty-four hours later, cells that migrated or invaded through the transwell membrane were fixed with 70% methanol, and then stained with 1% crystal violet (cat. no. AS1086; ASPEN Biotechnology). Subsequently, the migrated or invaded cells were observed using a fluorescence microscope and counted in five randomly selected fields.
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