Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose, ProClin 300.
Expression System:
HEK293 cells
Form:
Lyophilized powder
Sequence:
N-terminal His-Tag, Ser24~Ala92 (NP_002974.1)
Application Notes:
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 37C 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.
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