QCM ECMatrix Cell Invasion Assay, 24-well (8 µm), fluorimetric

Protocol tips

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	Add 10% fetal bovine serum) to the lower chamber. Cover plate and incubate for 24 - 72 hours at 37°C in a CO2 incubator (4-6% CO2).

Protocol tips
Add 10% fetal bovine serum) to the lower chamber. Cover plate and incubate for 24 - 72 hours at 37°C in a CO2 incubator (4-6% CO2).

Publication protocol

The QCM™ 24-well Fluorimetric Cell Invasion assay kit (ECM554; Chemicon) and the Migration kit (ECM509; Chemicon) were used to assess the migration and invasion properties of the miRNA-transfected SEOC cells. The assays were carried out according to the manufacturer’s protocol. Both kits used an insert polycarbonate membrane with an 8 μm pore size. The insert in the invasion kit was pre-coated with a thin layer of ECMatrix™ which occluded the membrane pores and blocked the migration of non-invasive cells. After 48 h transfection, cells were harvested and re-suspended in a serum-free medium. The upper chamber was filled with 300 μL cell suspension in triplicate in a 24-well plate, and 500 μl of complete medium was added to the bottom well. Following 24 h incubation, the cells that had not migrated were pipetted out from the upper surface of the inserts, and the cells that had migrated to the lower filters were detached using the cell detachment solution. Images of three random fields (×40 magnification) were captured from each well. The migrated and invaded cells were lysed and the relative invasion was determined by fluorescence with 480/520 nm filter set. The miRNA-transfected cells were normalized to their respective scrambled negative control groups.

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