Cultrex® Collagen IV Cell Invasion Assay

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	We used Cultrex 24-well collagen IV cell invasion assay kit (Trevigen, Gaithersburg, MD, USA). Transwell cell culture inserts with an 8-μm pore membrane were coated with collagen type IV. Cells (2 × 105 cells/insert) were seeded onto the upper chamber without FBS, and 500 μL of RPMI 1640 containing 10% FBS was loaded onto the bottom chamber. After 48 h of incubation, the cell dissociation solution was added onto the bottom chamber and incubated 1 h.

Protocol tips

We used Cultrex 24-well collagen IV cell invasion assay kit (Trevigen, Gaithersburg, MD, USA). Transwell cell culture inserts with an 8-μm pore membrane were coated with collagen type IV. Cells (2 × 105 cells/insert) were seeded onto the upper chamber without FBS, and 500 μL of RPMI 1640 containing 10% FBS was loaded onto the bottom chamber. After 48 h of incubation, the cell dissociation solution was added onto the bottom chamber and incubated 1 h.

Publication protocol

We used Cultrex 24-well collagen IV cell invasion assay kit (Trevigen, Gaithersburg, MD, USA). Transwell cell culture inserts with an 8-μm pore membrane were coated with collagen type IV. Cells (2 × 105 cells/insert) were seeded onto the upper chamber without FBS, and 500 μL of RPMI 1640 containing 10% FBS was loaded onto the bottom chamber. After 48 h of incubation, the cell dissociation solution was added onto the bottom chamber and incubated 1 h. The cell dissociation solution was quantitated with a microplate reader (Wako, Osaka, Japan) and the SoftMax Pro 5.1 software (Wako, Osaka, Japan). Each experiment was performed at least three times.

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