Mouse anti dsdna total ig elisa kit

Blood samples were collected by either retro-orbital eye bleeds under isoflurane anesthesia or cardiac puncture after euthanasia by CO₂ inhalation in accordance with Regeneron IACUC protocol. CBC with differential was performed on 20 μL of whole blood using Hemavet 950 (Drew Scientific Group) and clinical chemistry was completed on serum samples using ADVIA 1800 Chemistry System (Siemans Medical Solutions USA). ELISAs were performed on plasma samples using the following; Mouse IgG and IgM Rheumatoid Factor ELISA Kit (Shibayagi Co., Ltd), Mouse Anti-dsDNA Total Ig ELISA kit, Mouse ANA Total Ig ELISA kit, Mouse Anti-Sm Total Ig ELISA kit, Mouse Anti-Cardiolipin Total Ig ELISA kit (Alpha Diagnostic Intl.), and IgG and IgM mouse ELISA kit (Abcam) as per the manufacturer’s instructions. IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12total, IL-17, MCP-1, and TNF-α were measured in plasma samples using a Multi-Spot® 10plex electrochemiluminescence detection assay (Meso Scale Discovery) according to the manufacturer’s instructions.

Baseline antibody measurements were determined at weeks 9–10 of age and used to randomize animals to experimental groups using QuickCalcs tool from GraphPad (http://www.graphpad.com/quickcalcs/randomize2/). Each group consisted of a minimum of eight animals. Animals were orally administered vehicle, LAS191954 or prednisolone. All test solutions were analyzed after each period of administration to recheck stability and compound concentration. On weeks 2 and 4, blood was sampled by submandibular bleeding for antibody level determination. Samples from all time points were analyzed for anti-dsDNA levels and anti-desmoglein 3 antibody levels. Antibodies to dsDNA and Dsg3 were determined using the Mouse anti-dsDNA total Ig ELISA Kit (Alpha Diagnostic Cat#5110) and the Mouse Desmoglein 3 Antibody ELISA kit (MBL International Cat#7597), respectively.