Corning® BioCoat™ Tumor Invasion 24-well Plate

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Seed viable cells at 2×105/200μL after transfection in a serum free medium on theupper side of the well. - Add complete medium on the lower side.	- Treat upper chamber with Matrigel matrix membrane after 48h of culture.	- Stain with 0.2% w/v crystal violet before counting cells.

Upstream tips
- Seed viable cells at 2×105/200μL after transfection in a serum free medium on theupper side of the well. - Add complete medium on the lower side.
Protocol tips
- Treat upper chamber with Matrigel matrix membrane after 48h of culture.
Downstream tips
- Stain with 0.2% w/v crystal violet before counting cells.

Publication protocol

The invasion ability of the cells with or without treatment was tested by using the BD BioCoat Tumor Invasion Assay System (BD Biosciences, Bedford, MA, USA) according to the manufacturer’s protocol. Briefly, osteosarcoma cells that were untransfected or transfected with the respective control vector or Notch‐1 plasmids were treated with curcumin at various concentrations for 48 h. Next, viable cells (2 × 105/200 μL, confirmed by trypan blue exclusion) were seeded in serum‐free medium onto the upper chamber, while complete medium was added in the lower compartment as a chemoattractant. After 48 h of incubation, the cells in the upper chamber were removed and the cells that invaded through the Matrigel matrix membrane were fixed with 70% ethanol and stained with 0.2% w/v crystal violet before being counted using the Olympus Optical microscope (Melville, NY, USA). Invaded cells from six random microscopic fields (100 × magnification) were enumerated and all experiments were performed in triplicate.

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Manufacturer protocol

Download the product protocol from Corning for Corning® BioCoat™ Tumor Invasion 24-well Plate below.

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