Corning® BioCoat™ Matrigel® Invasion Chamber with 8.0 µm PET Membrane in four 6-well Plates

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Plate should be stored at -20C in the original packaging.	Incubate the Corning BioCoat™ Matrigel Invasion Chambers for 24 hours in a humidified tissue culture incubator, at 37C, 5% CO2 atmosphere. Scrubbing must be accomplished quickly to avoid drying of the cells adhering to the bottom surface of the membrane Stain cells with 0.05% crystal violet for 15 minutes

Upstream tips
Plate should be stored at -20C in the original packaging.
Protocol tips
Incubate the Corning BioCoat™ Matrigel Invasion Chambers for 24 hours in a humidified tissue culture incubator, at 37C, 5% CO2 atmosphere. Scrubbing must be accomplished quickly to avoid drying of the cells adhering to the bottom surface of the membrane Stain cells with 0.05% crystal violet for 15 minutes

Publication protocol

For invasion assays, cells were plated (105 cells per chamber) onto BD BioCoat Matrigel invasion chambers (BD Biosciences, San Josè, CA). In the upper chamber, the medium was supplemented with 2% heat-inactivated FBS. In the lower chamber, 20% FBS was used as a chemoattractant. PCPA was added in the upper chamber. After 24 hours, the medium was removed and the chambers were washed twice with phosphate-buffered saline (PBS); non-invading cells were removed from the upper surface of the membrane by scrubbing with a cotton-tipped swab; invading cells were fixed with 3.7% formaldehyde in PBS for 2 minutes, washed twice with PBS, permeabilized with methanol for 20 minutes, washed twice with PBS, stained with 0.05% crystal violet for 15 minutes, and washed twice with PBS. Ten fields for each chamber were photographed using a digital camera mounted on an inverted microscope (magnification, × 10), and invading cells were counted in each field. Experiments were carried out in duplicate and repeated twice.

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Manufacturer protocol

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