CytoSelect™ 24-Well Wound Healing Assay

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- During washing step, cells are washed once with 1 ml of growth medium and twice with 1 ml of medium containing 0.5% FCS.	- Cells are assessed after 2, 4, 8 and 24 hours by photography.

Protocol tips
- During washing step, cells are washed once with 1 ml of growth medium and twice with 1 ml of medium containing 0.5% FCS.
Downstream tips
- Cells are assessed after 2, 4, 8 and 24 hours by photography.

Publication protocol

"The in vitro cell migration was measured by the scratch assay method described by Liang [22], with some modifications. After trypsinization, cells were plated in 2 ml of medium in poly-L-lysine coated 12-wells plates and placed in a 37°C, 5% CO2 incubator. Cells were grown until a confluent monolayer was formed. With a 200ul pipet tip a scratch was made in the cell monolayer. The debris was removed by washing the cells once with 1 ml of growth medium and twice with 1 ml of medium containing 0.5% FCS. Hereafter, 2 ml of medium containing 0.5% FCS and the different compounds of interest were added. The ability of cells to migrate into the scratch area was assessed after 2, 4, 8 and 24 hours by comparing the 0- and 2,4,8 and 24-hour photomicrographs (Zeiss, Axiovert 40c, x50 magnification) of 4 fixed points along the scratch area. The percentage of non-recovered scratch area was calculated by dividing the non-recovered area after 2, 4, 8 and 24-hours by the initial scratch area (t=0) using the image software (http://rsb.info.nih.gov/ij/).

The scratch assay method was compared with a wound healing assay using the Cytoselect ™Wound Healing Assay Kit (Cell Biolabs, Inc., San Diego, USA). Briefly, after trypsinization cells were seeded in 1 ml of growth medium into 24-wells tissue culture plates containing inserts and incubated until a monolayer of cells was formed. After removing the inserts from the wells, the cells were treated according to the scratch assay method as described above"

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Manufacturer protocol

Download the product protocol from Cell Biolabs for CytoSelect™ 24-Well Wound Healing Assay below.

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