Biocoat 8.0 μm pet membrane 24 well cell culture inserts

0.3 ml serum free media containing 0.3×10⁵ GIST-T1, GIST430, RMS176 or LMS04 cells were plated for invasion assays in BD BioCoat^™ Matrigel Invasion Chambers (BD Biosciences, Catalog # 354480) and for migration assays in BD BioCoat^™ 8.0 μm PET Membrane 24-well Cell Culture Inserts (BD Biosciences, Catalog # 354578). The wells were fed with 0.5ml RPMI Media 1640 (Invitrogen) containing 15% FBS and incubated in a humidified incubator, at 37°C, 5% CO₂ for 144 hours (GIST-T1), 72 hours (GIST430) or 24 hours (RMS176, LMS04, and HT-1080). The media from the inside of the insert was aspirated, and the interiors of the inserts were gently swabbed to remove non-invasive or non-migratory cells. Inserts were transferred to new wells containing 400 μl Cell Stain Solution (Cell Biolabs, Inc) and incubated for 10 minutes at room temperature, then rinsed two times in a beaker of water. Then, the inserts were air dried and transferred to new wells containing 200 μl Extraction Solution (Cell Biolabs, Inc) and quantified at OD 560nm in a microplate reader.