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:
HEK293 cells
Form:
Lyophilized powder
Sequence:
His-Tag and Fc-Tag, Thr678~Arg751 (NP_001726.2)
Application Notes:
Complement Component 5a (C5a) is a component of the complement system which plays a key role in promoting migration and adherence of neutrophils and monocytes to vessel walls. C5a has been proven to be able to induce chemotactic migration of THP-1 cells. Therefore, chemotaxis assay used 24-well microchemotaxis system was undertaken to detect the chemotactic effect of C5a on the human monocytic cell line THP-1. Briefly, THP-1 cells were seeded into the upper chambers (100 µl cell suspension, 106 cells/ml in RPMI-1640 with 0.5% FBS) and C5a (50 and 100 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 2h, 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). C5a is able to induce migration of THP-1 cells.
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