The Enzyme-Linked Immunosorbent Assay (ELISA) ( Abcam, UK) was applied to determine the serum levels of CX3CL1, CXCL5, CXCL7, CXCL9, CXCL11 and CCL28 using specific enzyme immune assay test kits following the manufacturer's protocol.
Microplates were coated with human antibodies specific for antigens of the different chemokines. All immunoenzymatic techniques were based on measuring the absorbance after a peroxidation reaction at 450 nm.
Between the reaction steps, the plates were washed manually. The absorbance was measured using a microplate reader (MR-96A).
The concentrations were extrapolated from an obtained standard curve.