Cultrex BME Cell Invasion Assay, 96 well

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Coat upper chamber with 0.5 X BME solution at 37°C in a CO2 incubator for 4 hours	Incubate bottom chamber at 37C in CO2 incubator for 24 hours

Upstream tips
Coat upper chamber with 0.5 X BME solution at 37°C in a CO2 incubator for 4 hours
Protocol tips
Incubate bottom chamber at 37C in CO2 incubator for 24 hours

Publication protocol

One day prior to beginning the assay, the AGS cells was starved in a serum-free medium. Invasion assay was performed using the Culturex 96 well BME invasion assay kit. Briefly, top chamber was coated with 0.5 X BME solution at 37°C in a CO2 incubator for 4 hours and then plated with 5 × 104 cells in serum-free DMEM/F12 medium with 10mM metformin. Bottom chamber was filled with medium containing FBS and incubated at 37°C with 5% CO2 for 24 hours. After washing the chambers, diluted Calcein–AM stock solution was added into the bottom chamber and photographed by fluorescence microscope. The total numbers of invaded cells were counted by imageJ software (NCBI). All experiments were independently repeated at least three times.

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