Active Stromal Cell Derived Factor 1 (SDF1), RPU54874
SDF1 (stromal cell-derived factor 1), also known as C-X-C motif chemokine 12, is a chemokine protein that has chemotaxis active on T-lymphocytes and monocytes. It is thought that SDF1 stimulates migration of monocytes and T-lymphocytes through its receptors, CXCR4 and ACKR3; thus, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of SDF1 on the human monocytic cell line THP-1. Briefly, THP-1 cells were seeded into the upper chambers (100uL cell suspension,106cells/mL in RPMI 1640 with 0.5% FBS) and SDF1 (10ng/mL, 30ng/mL and 60ng/mL diluted separately in serum free RPMI 1640) was added in lower chamber with a polycarbonate filter (8µm pore size) used to separate the Two compartments. After incubation at 37℃ with 5% CO2 for 3h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×100) and the number of migrated cells were counted at high magnification (×400) randomly (five fields for each filter). Result shows SDF1 is able to induce migration of THP-1 cells. The migrated THP-1 cells in low chamber at low magnification (×100) were shown in Figure 1. Five fields of each chamber were randomly chosen, and the migrated cells were counted at high magnification (×400). Statistical results were shown in Figure 2. The optimum chemotaxis of SDF1 occurs at 10ng/mL.
Figure 3. SDS-PAGE
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