Human baff instant elisa

Serum samples from patients and controls were maintained at −80°C until cytokine determinations. IFNα, IL-17A, IL-10, IL-6, CCL3 (MIP-1α), and CCL5 (RANTES) concentrations were quantified by Cytometric Bead Arrays Flex Set by using a FACS Canto II flow cytometer (BD) and following the manufacturer's instructions. For IL-10 and IL-6, an Enhanced Sensitivity Flex Set was used. Finally, ELISA kits were used for the quantification of TNFα, CCL2 (MCP-1), CXCL10 (IP-10) (Mini kit, PeproTech), IFNγ (OptEIA kit, BD) and BLyS (Human BAFF Instant ELISA, eBioscience) following the manufacturer's instructions. The lower limits of detection were 1.25 pg/ml for IFNα, 0.30 pg/ml for IL-17A, 0.014 pg/ml for IL-10 and IL-6, 0.20 pg/ml for CCL3, 0.002 pg/ml for CCL5, 3.90 pg/ml for TNFα, 8.00 pg/ml for CCL2, 16.00 pg/ml for CXCL10, 0.58 pg/ml for IFNγ, and 0.13 ng/ml for BLyS.

Serum levels of circulating cytokines were analyzed by commercial immunoassays following manufacturer instructions as follows: sBLyS [Human BAFF instant ELISA, eBioscience, detection limit (d.l.) = 0.13 ng/ml], sBCMA (TNFRSF17 ELISA kit, Raybiotech, d.l. = 0.025 ng/ml), sTACI (Human TACI ELISA kit, Raybiotech, d.l. = 0.32 pg/ml), sAPRIL (Human APRIL Platinum ELISA, eBioscience, d.l. = 0.19 ng/ml), sBLyS-R (Human TNFRSF13C, abcam, d.l. = 0.78 pg/ml), IL-10 (Human IL-10 ELISA high sentivity, eBioscience, d.l. = 0.19 pg/ml), IL-37 (Human IL-37/IL1F7 ELISA, BosterBio, d.l. = 4.8 pg/ml), TNFα (Human TNFα Mini EDK, Peprotech, d.l. = 3.9 pg/ml) and IFNg (IFNg OptEIA kit, BD Bioscience, d.l. = 0.58 pg/ml). Levels of IFNα, MIP1α, IL-8 and GM-CSF were quantified using a Cytometric Bead Array Flex Set (BD) in a FACS Canto II flow cytometer. The detection limits were 1.25, 0.2, 1.2, and 0.2 pg/ml, respectively.

Serum samples from SLE patients and HC were collected and maintained at −80 °C until cytokine determination was carried out. IL-1β, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-17 A, IFNα, VEGF and GM-CSF amounts were analyzed by Cytometric Bead Arrays Flex Set using a FACS Canto II flow cytometer (BD Biosciences). For IL-1β, IL-6, IL-10 and IL-12p70, an Enhanced Sensitivity Flex Set was needed. ELISA kits were used for the quantification of TNFα, leptin, resistin (Mini EDK kit, PeproTech), BLyS (Human BAFF Instant ELISA, eBioscience) and IFNγ (OptEIA kit, BD) following the manufacturer’s instructions. The lower limits of detection were 48.4 fg/ml for IL-1β, 1.4 pg/ml for IL-4, 68.4 fg/ml for IL-6, 1.2 pg/ml for IL-8, 13.7 fg/ml for IL-10, 12.6 fg/ml for IL- 12p70, 0.3 pg/ml for IL-17 A, 1.25 pg/ml for IFNα, 4.5 pg/ml for VEGF, 0.2 pg/ml for GM-CSF, 3.9 pg/ml for TNFα, 63 pg/ml for leptin, 24 pg/ml for resistin, 130 pg/ml for BLyS and 0.58 pg/ml for IFNγ.