5 µm Chemotaxis Assays, 24-Well Format

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Store all components at 4ºC.	Incubate for 24 hours in a cell culture incubator at 37C. Retain the medium in the 24-well migration plate that contains chemoattractant(s) and cells that migrated through the membrane and into the medium	Read fluorescence with a fluorescence plate reader at 480 nm/520 nm.

Upstream tips
Store all components at 4ºC.
Protocol tips
Incubate for 24 hours in a cell culture incubator at 37C. Retain the medium in the 24-well migration plate that contains chemoattractant(s) and cells that migrated through the membrane and into the medium
Downstream tips
Read fluorescence with a fluorescence plate reader at 480 nm/520 nm.

Publication protocol

The transwell cell migration assay was performed using a Fluorometric Cell Migration Assay kit with polycarbonate membrane inserts (5-μm pore size; Cell Biolabs, San Diego, CA, USA). Cells were serum-starved overnight in DMEM prior to initiation of the experiment. Cells were then incubated at 37°C for 24 hours to allow cell migration through the membrane. Migratory cells were detached from the underside of the membrane and subsequently lysed and detected by CyQuant GR dye (Invitrogen). Fluorescence measurement was performed in a FluoStar Optima fluorescence plate reader with a 485 / 520 nm filter set.

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