96-Well Cell Invasion Assay, Basement Membrane

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Incubate at 37C in a CO2 incubator for 12 hours	Read at 480 nm/520 nm

Protocol tips
Incubate at 37C in a CO2 incubator for 12 hours
Downstream tips
Read at 480 nm/520 nm

Publication protocol

The invasion assay was carried out following the instructions of Cultrex 24-well Transwell BME cell invasion assay (Trevigen). Briefly, a 24-well unit with 8 µm polycarbonate nucleopore filters (Corning) evenly coated with 100 µl basement membrane extract coating solution (Trevigen) at 37°C for 4 h. Control A549 cells (2×105) expressing GFP and RAW 264.7 macrophages (1×105) in serum free medium were placed in the upper compartment, and 50% RPMI 1640 plus 50% DMEM medium supplied with 0.5% FBS was added to the lower compartment. After 24 h incubation, cells that had not invaded were removed with a cotton swab. Cells that had invaded the lower surface of the membrane were observed under a fluorescence microscope. Invaded cells in three randomly selected fields were counted.

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