Human MIP1 alpha protein, His tag (active)

MIP-1a (macrophage inflammatory protein 1-alpha) also known as Chemokine (C-C motief) ligand 3 (CCL3), is a cytokine belonging to the CC chemokine family that is involved in the recruitment and activation of macrophages, monocytes and neutrophils. In this case, chemotaxis assay used 24-well microchemotaxis system was undertaken to evaluate the chemotactic effect of MIP-1a on the human monocytic cell line THP-1. Briefly, THP-1 cells were seeded into the upper chambers (100 μl cell suspension, 1x10⁶ cells/ml in RPMI-1640 with 0. 5% FBS) and MIP-1a (100 ng/ml, diluted 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 37ºC with 5% CO₂ for 5h, the filter was removed, then cells in low chamber were observed by inverted microscope at low magnification (×40) and the number of migrated cells were counted at high magnification (×400) randomly (five fields for each filter). By counting migrated cells in low chamber at high magnification (×400) randomly. A mean of 41. 2 THP-1 cells/field migrated towards serum free RPMI-1640 medium with 100 ng/ml MIP-1a, while only 3. 6 THP-1 cells/field migrated towards serum free RPMI-1640 medium.