Reconstitute with 20mM Tris (pH8.0) and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris (pH8.0), 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose, ProClin 300.
Expression System:
E. coli
Formulierung:
Lyophilized powder
Sequenz:
N-terminal His-Tag, Gly22~Ser98 (NP_006265.1)
Anwendungsbeschreibung:
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 37C 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.
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