Chemotaxis assays were performed as described previously 53 (link). 5×105 enriched CD4 splenic T cells were incubated for 1 h with 1× DMEM containing 0.5% fatty acid–free BSA (EMD Biosciences), 5% antibiotics, L-glutamine (Cellgro), and Hepes. Cells were then allowed to migrate through 5-μm-pore–sized transwells (Corning) toward soluble CXCL12, CXLC13 (R&D Systems) or media alone, for 3 h at 37°C. Cells were collected, stained, and resuspended in 45 μl of staining buffer and analyzed by flow cytometry.
5 μm pore sized transwells
Manufactured by Corning
The 5-μm-pore–sized transwells are a laboratory equipment product designed for cell culture applications. They feature a porous membrane with a pore size of 5 micrometers, allowing for the exchange of nutrients, gases, and signaling molecules between the upper and lower chambers of the transwell system.
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2 protocols using 5 μm pore sized transwells
1
Chemotaxis Assay for CD4+ T Cells
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Chemotaxis Assay for CD4+ T Cells
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Chemotaxis assays were performed as described previously 53 (link). 5×105 enriched CD4 splenic T cells were incubated for 1 h with 1× DMEM containing 0.5% fatty acid–free BSA (EMD Biosciences), 5% antibiotics, L-glutamine (Cellgro), and Hepes. Cells were then allowed to migrate through 5-μm-pore–sized transwells (Corning) toward soluble CXCL12, CXLC13 (R&D Systems) or media alone, for 3 h at 37°C. Cells were collected, stained, and resuspended in 45 μl of staining buffer and analyzed by flow cytometry.
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