Human complement c5a elisa kit

The levels of anaphylatoxins C3a and C5a were measured as an indicator of complement activation. Cells were incubated with 1:8 diluted normal human serum (NHS) (Complement Technology) which was used as a source of complement. After 4 h of incubation, the conditioned medium was collected and measured for C3a and C5a levels using Complement C3a Human ELISA Kit (Thermo Fisher) and Human Complement C5a ELISA Kit (Abcam), respectively. All measurements were performed in triplicates.

C3a desArg, C5a, Bb and sC5b‐9 were detected by sandwich enzyme‐linked immunosorbent assay (ELISA): complement C3a desArg human ELISA kit (cat. no. ab133037; Abcam, Cambridge, MA, USA), human complement C5a ELISA kit (cat. no. ab193695; Abcam, Cambridge, MA, USA), MicroVue Bb plus fragment EIA (cat. no. A027; Quidel Crop, Pathway diagnostics Ltd, Dorking, UK) and MicroVue SC5b‐9 Plus EIA (cat. no. A029; Quidel Crop, Pathway diagnostics Ltd, Dorking, UK). Absorbance was measured at 450 nm (NS‐100 Nano Scan Microplate Reader), and concentrations were determined using the commercial standard curves provided with the kits.