Human MIP3 beta protein, His tag (active)

Macrophage Inflammatory Protein 3 Beta (MIP3b) is a small cytokine belonging to the CC chemokine family that is also known as EBI1 ligand chemokine (ELC) and Chemokine C-C motif ligand 19 (CCL19). This chemokine elicits its effects on its target cells by binding to the chemokine receptor chemokine receptor CCR7. It attracts certain cells of the immune system, including dendritic cells and antigen-engaged B cells, CCR7 central-memory T-Cells. Thus, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of recombinant human MIP3b on the Jurkat cell line. Briefly, Jurkat cells were seeded into the upper chambers (150 μl cell suspension,10⁶ cells/ml in RPMI-1640 with FBS free) and MIP3b (0.01, 0.1, 1, 10, 100, and 1000 ng/ml diluted separately in serum free RPMI-1640) was added in lower chamber with a polycarbonate filter (8 μm pore size) used to separate the two compartments. After incubation at 37ºC 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). MIP3b is able to induce migration of Jurkat cells, and the migrated cells were counted at high magnification (×400). The optimum chemotaxis of recombinant human MIP3b occurs at 0.1-1 ng/ml.