Qcmtm 24 well cell invasion assay kit

In vitro cell invasion assays were performed using a modified Boyden chamber assay. Cells were added to the upper chamber of QCMTM 24-Well Cell Invasion Assay Kit (Millipore) inserts. Medium containing serum (10%) was utilized as the chemoattractant in the lower chamber. After 24 hours of incubation, cells that had invaded to the lower surface of the collagen-coated membrane were analyzed according to the manufacturer's instructions.
Cell extracts were prepared 24 hours after transfection, and luciferase activity was measured using the Dual-Luciferase Reporter Assay System (Promega). All experiments were performed three times in triplicate.

4 × 10⁵ cells per well were plated into 6-well plates and allowed to grow for 24 h followed by treatment. Twenty-four hours later, a scratch was introduced in each well using a pipette tip. After washing with PBS twice, serum-free medium was used for cell culture, and cells were allowed to grow for a further 24 and 48 h. Microphotographs were taken under a phase-contrast microscope.
Invasion assays were performed using a QCM^TM 24-Well Cell Invasion Assay Kit with 8 μm membrane (Millipore, Bedford, MA, USA). Cells were seeded at a density of 5 × 10⁴/chamber well in triplicate with serum-free medium and were allowed to invade through the matrigel for 48 h. Cells that had invaded to the lower surface of the membrane were fixed with methanol and stained with 0.5% crystal violet solution, then were photographed and counted by inverted microscope.