Human ccl3 mip 1 alpha quantikine elisa kit

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.

Baseline blood specimens were collected from storage in July 2012. The sera were snap frozen and stored at −70 °C until further analysis and underwent only one freeze-thawing cycle. Baseline serum levels of CCL3/MIP-1α, CCL5/RANTES and CCL18/PARC were determined by commercially available ELISA kits from R&D systems (Abingdon, United Kingdom; Human CCL3/MIP-1 alpha Quantikine ELISA Kit, ≤8.9 % intra-assay variation, Human CCL 5/RANTES DuoSet and Human CCL18/PARC DuoSet). CCL3 ELISA was performed according to the manufacturer’s instructions. For CCL5 measurements, 0.75 μg/ml capture antibody and 10 ng/ml detection antibody were used, for CCL18 we used 1.5 μg/ml capture antibody and 200 ng/ml detection antibody.