For the neutrophil migration assay, Hoxb8 neutrophils were washed with the migration medium RPMI-1640 containing 2.5mM HEPES and 0.1% BSA and tested for migration in 96-well microchamber using a 5 μm-pore size, polycarbonate filter (Boyden Chamber, Neuro Probe Inc., Gaithersburg, MD, USA). The lower wells of the Boyden Chambers were filled with the migration medium supplemented with either PBS or 10nM murine recombinant chemokine (C-C motif) ligand 3 (Ccl3) (R&D). In the upper wells, 5 × 104 Hoxb8 neutrophils were added and the chamber were incubated for 90 minutes at 37 °C and 5% CO2. After incubation, nonmigrating cells in the upper chamber were removed using a cotton-tipped applicator. Cells that had migrated to the bottom side of the membrane were fixed in 70% ethanol and stained with DAPI before mounting onto poly-lysine coated slides. Each condition was performed in duplicate. The number of migrated neutrophils counted in five random fields was analyzed using Fiji (Version 2.1.0/1.53h).
Murine recombinant chemokine c c motif ligand 3 ccl3
Manufactured by R&D Systems
Murine recombinant chemokine (C-C motif) ligand 3 (Ccl3) is a lab equipment product that functions as a chemokine. Chemokines are a family of small cytokines that are important for the recruitment and activation of immune cells.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using murine recombinant chemokine c c motif ligand 3 ccl3
1
Hoxb8 Neutrophil Migration Assay
2
Hoxb8 Neutrophil Migration Assay
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For the neutrophil migration assay, Hoxb8 neutrophils were washed with the migration medium RPMI-1640 containing 2.5mM HEPES and 0.1% BSA and tested for migration in 96-well microchamber using a 5 μm-pore size, polycarbonate filter (Boyden Chamber, Neuro Probe Inc., Gaithersburg, MD, USA). The lower wells of the Boyden Chambers were filled with the migration medium supplemented with either PBS or 10nM murine recombinant chemokine (C-C motif) ligand 3 (Ccl3) (R&D). In the upper wells, 5 × 104 Hoxb8 neutrophils were added and the chamber were incubated for 90 minutes at 37 °C and 5% CO2. After incubation, nonmigrating cells in the upper chamber were removed using a cotton-tipped applicator. Cells that had migrated to the bottom side of the membrane were fixed in 70% ethanol and stained with DAPI before mounting onto poly-lysine coated slides. Each condition was performed in duplicate. The number of migrated neutrophils counted in five random fields was analyzed using Fiji (Version 2.1.0/1.53h).
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