Calcium Flux/Mobilization Assay Protocol
Introduction
To test the migration ability of cells in response to the simulation by chemokine. The EC50 of each chemokine will depend on the cell line and receptor. For example, to test the migration of CXCR4 expressed in Jurkat cells in response to SDF-1a, use 0.003, 0.012, 0.048, 0.019, 0.75. 3.0 nM SDF-1a.
Protocol
Introduction
To test the migration ability of cells in response to the simulation by chemokine. The EC50 of each chemokine will depend on the cell line and receptor. For example, to test the migration of CXCR4 expressed in Jurkat cells in response to SDF-1a, use 0.003, 0.012, 0.048, 0.019, 0.75. 3.0 nM SDF-1a.
Protocol
- Prepare a chemokine concentration gradient in 6 wells of a Transwell plate containing 600ul of growth media.
- Add 100ul of a cell suspension at 2.5 million cells/ml to the transwell inserts and place each in the bottom chamber containing the media and chemokine. Incubate for 2 hours at 37C and 5% CO2.
- The cells that migrate through the membrane pores to the bottom chamber are counted by flow cytometer.
