Human cx3cl1 fractalkine quantikine elisa kit

CX3CL1 was quantified using the Human CX3CL1/Fractalkine Quantikine ELISA Kit (R&D Systems) according to manufacturer’s instructions in the same subset of patients, from whom an expression marker analysis was performed by flow cytometry (FC) (n = 34). Optical density was measured on a Promega GloMax Discover microplate reader (450 nm, reference 620 nm).

The cell supernatants, rat sera, and rat lung tissue homogenates were harvested and stored at −80°C. The levels of FKN were measured by the human CX3CL1/fractalkine Quantikine ELISA kit (DCX310, R&D Systems Inc., Minneapolis, USA) and rat fractalkine (CX3CL1) ELISA kit (ab100761, Abcam Ltd., Cambridge, United Kingdom) according to the manufacturer's instructions. The levels of LBP were measured by the human LBP DuoSet ELISA kit (DY008, R&D Systems Inc., Minneapolis, USA) and rat LBP ELISA kit (CSB-E11184r, Cusabio, Wuhan, China) according to the manufacturer's instructions. The BALF supernatants and rat lung tissue homogenates were harvested and stored at −80°C. The levels of myeloperoxidase activity were measured by the rat MPO ELISA kit (HK105-01, Hycult Biotech, USA) according to the manufacturer's instructions.

Plasma was obtained from whole blood, drawn in EDTA tube, by centrifuging at 400xg, for 10 min at room temperature. Plasma samples from IgAN patients and HC were then analyzed with enzyme-linked immunosorbent assay (ELISA) to determine the concentration of the following factors; monocyte chemoattractant protein (MCP-1) (Human CCL2/MCP-1 Quantikine ELISA Kit, R&D Systems, USA), soluble CD14 (Human CD14 Quantikine ELISA Kit, R&D Systems), soluble CD40L (Human CD40 Ligand/TNFSF5 Quantikine ELISA Kit, R&D Systems), B cell activating factor (BAFF) (Human BAFF Quantikine ELISA Kit, R&D Systems), IL-6 (Human IL-6 Quantikine HS ELISA Kit, R&D Systems), Fractalkine (Human CX3CL1/Fractalkine Quantikine ELISA Kit, R&D Systems) and MIP-1 (Human CCL3/MIP-1 alpha Quantikine ELISA Kit, R&D Systems). Catalog number for ELISA kits are shown in S2 Table. The samples were assayed in duplicates and the average values were considered for further analysis. The optical density (OD) in blank well was subtracted from the OD values in samples before calculation of concentrations.