8 µm Chemotaxis Assays, 24-Well Format

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	Incubate for 4-6 hours in a cell culture incubator

Protocol tips
Incubate for 4-6 hours in a cell culture incubator

Publication protocol

The experiment was carried out using the CytoSelect cell migration assay kit (Cell Biolabs, Inc., San Diego, CA) containing polycarbonate membrane inserts (8 μm pore size) in a 24-well plate. In brief, appropriate amount of cells were suspended in serum-free medium and added to the insert. The lower well was loaded with routine medium (with 10% FBS). After an incubation of 4-6 hours at 37°C in a humidified incubator with 5% CO2, non-migratory cells on the upper surface of the membrane were gently removed by wiping with a cotton swab. The cells that migrated through the pores and to the underside of the membrane were fixed, stained and photographed under a microscope. The number of cells was counted and averaged using three randomly selected areas per membrane. Doxycycline (0.8 µg/mL) was added to medium/cells over the time course of the desired experimental groups.

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