24-Well Cell Invasion Assays, Basement Membrane

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Incubate for 48 hours at 37ºC in 5% CO2 atmosphere	Measure absorbance at 560 nm

Protocol tips
Incubate for 48 hours at 37ºC in 5% CO2 atmosphere
Downstream tips
Measure absorbance at 560 nm

Publication protocol

Invasion assays were conducted using the CytoSelect 24-Well Cell Invasion Assay Kit (Cell Biolabs, San Diego, CA, USA). Briefly, 300 μL of HCT116 and HT29 cells (1 × 105cells/ml) in serum-free medium was plated into the CytoSelect basement membrane chamber, respectively, and 500 μL of 10% FBS-containing RPMI was added to the lower well of the invasion plate; both upper and lower chambers contained the 10 uM IWR-1. The chambers were then incubated for 48 h at 37°C in a 5% CO2 atmosphere, non-migratory cells were removed, and migrated cells were stained, dissociated from the membrane, and their absorbance was measured at 560 nm using the microplate reader (model 680; Bio-Rad, Hercules, CA, USA) [65].

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