3 µm Chemotaxis Assays, 96-Well Format

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Cover the plate and transfer to a cell culture incubator for 2 hours.	Read the fluorescence with a fluorescence plate reader at 480 nm/520 nm.

Protocol tips
Cover the plate and transfer to a cell culture incubator for 2 hours.
Downstream tips
Read the fluorescence with a fluorescence plate reader at 480 nm/520 nm.

Publication protocol

Chemotaxis was assayed using the CytoSelect 96-well Cell Migration Assay Kit (3 μm pore, Cell Biolabs, INC.) 5 × 105 Jurkat cells were suspended in serum free RPMI media and loaded into the upper chamber of the transwell. The bottom chamber contained media and 40 ng/ml SDF-1α (Peprotech Inc., Rocky Hill, NJ). Cells were irradiated with visible light (5–10 mW cm−2) or a blue light photodiode array (470 nm, 10 mW cm−2) and then incubated for 120 min at 37 °C in a CO2 incubator. Migrated cells were counted manually using a hemocytometer. Cell motility assays were performed on fibronectin (2 μg/ml) coated coverslips at 30–32 °C. Individual cells were tracked using CellTrack software50. We analyzed cells for 4 min before and after blue-light irradiation.

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