24-Well Cell Invasion Assays, Basement Membrane

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Store all components at 4ºC and before use allow the invasion chamber plate to warm up at room temperature for 10 minutes.	Incubate for 72 hours at 37ºC in 5% CO2 atmosphere.	Measure the OD 570nm in a plate reader

Upstream tips
Store all components at 4ºC and before use allow the invasion chamber plate to warm up at room temperature for 10 minutes.
Protocol tips
Incubate for 72 hours at 37ºC in 5% CO2 atmosphere.
Downstream tips
Measure the OD 570nm in a plate reader

Publication protocol

In vitro cell invasion assay was performed using a basement membrane-coated CytoSelect™ 24-well Cell Invasion Assay Kit (8 mm pores, Colorimetric Format, Cell Biolabs, San Diego, CA, US) according to the manufacturer’s instruction. Briefly, CD10- or mock-A375 cells were serum-starved for 24 h. Next, the cells suspended with serum-free media (3 X 105 cells/well) were seeded in the upper chamber separated from a lower chamber filled with media containing 10% FBS and incubated for 72 h. After noninvasive cells were removed, the invasive cells were stained and quantified by measuring the absorbance at 570 nm using a microplate reader (iMark™ Microplate Absorbance Reader, Bio-Rad Laboratories). All assays were carried out 3 times in triplicates.

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