Cultrex® BME Cell Invasion Assay

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 50,000 cells	Incubate at 37oC in CO2 incubator for 24 hours	Read assay chamber solutions (bottom) at 485 nm excitation, 520 nm emission

Upstream tips
Seed 50,000 cells
Protocol tips
Incubate at 37oC in CO2 incubator for 24 hours
Downstream tips
Read assay chamber solutions (bottom) at 485 nm excitation, 520 nm emission

Publication protocol

Experiments were performed using a Cultrex 96 Well Basement Membrane Extract (BME) Cell Invasion Assay kit (Trevigen) and medium containing 2.5 µg/ml Mitomycin C. For Nodal induction, ~ 50,000 T47D cells in serum free medium containing 0, or 39 nM (500 ng/ml) Nodal were seeded in the top chamber of the BME coated Boyden chamber plate. Serum free medium containing 0.2 units/ml bovine insulin was added to the bottom chamber. For Cerberus inhibition, about 10,000 MCF-7, Hs578t, BT-549, and MDA-MB-231 cells in serum free medium were seeded in the top chamber of the BME coated Boyden chamber plate. Complete medium was added to the bottom chamber. Top and bottom chamber contained 0 nM, 17.8 nM, or 178 nM Cerberus-Fc and 2.5 µg/ml Mitomycin C. Cell invasion was determined after 24 h by monitoring Calcein-AM fluorescence in the bottom chamber.

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