Mouse baff blys tnfsf13b quantikine elisa kit

The levels of BAFF in media, imiquimod, or ODN 2395 stimulated cell culture supernatants were analyzed using the Mouse BAFF/BLyS/TNFSF13B Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA). The levels of IFNα in serum and splenic leukocyte culture supernatants were determined with eBioscience Mouse IFN alpha Platinum ELISA kit (eBioscience). The ELISAs were performed per the manufacturers’ protocols. Ciraplex® Chemiluminescent Assay kits (Aushon Biosystem, Billerica, MA, USA) were used to quantify the levels of IFN-γ, IL-1β, IL-2, IL-6, IL-10, and TNFα in serum and cell culture supernatants per the manufacturer’s instructions^{49 (link),65 (link)}. The images of chemiluminescent array plates were captured with Cirascan image system (Aushon) and the image data was processed with Cirasoft software. All the assays were performed with two technical replicates for each sample.

At necropsy blood was collected from mice by cardiac puncture and placed into EDTA-tubes to prevent coagulation. Blood Samples were centrifuged and plasma was collected and frozen at −80°C until used. Cytokines and chemokines were measured using the V-PLEX Plus Pro-inflammatory Panel1 Mouse Kit from MSD (Rockville, MD). Plasma BAFF concentrations were measured using the Mouse BAFF/BLyS/TNFSF13B Quantikine ELISA Kit from R&D Systems (Minneapolis, MN) following the manufacturers protocol. Total IgG was quantified using IgG (Total) Mouse Uncoated ELISA kit (Invitrogen) following the manufacturers protocol. Anti-ds-DNA IgG present in the kidney was quantified using Mouse Anti-dsDNA IgG Antibody Assay Kit (Chondrex, inc.) following the manufacturers protocol. Plasma creatinine was measured using the Creatinine (serum) Colorimetric Assay Kit (Cayman Chemical; Ann Arbor, MI).

BAFF serum levels were assessed using the Mouse BAFF/BLyS/TNFSF13B Quantikine ELISA Kit (R&D Systems), and concentrations obtained by comparing to a standard curve.