Mouse anti dsdna igg elisa kit

Serum anti-dsDNA titers were measured using mouse anti-dsDNA IgG ELISA kit (Alpha Diagnostic International Inc.) according to the manufacturer’s instruction. Proteinuria was measured using Albustix Reagent Strips (SIEMENS) according to the manufacturer’s instruction.

For HID feeding and SRBC immunization, 3-week-old B6 mice were pretreated with HID for 5 weeks and then were i.p. immunized with 5% SRBCs in Alsever’s solution. To analyze the role of miR-21 on TD humoral response, 8-week-old miR-21 cKO mice and WT controls were injected i.p. with 5% SRBCs in Alsever’s solution. After 7–14 days of immunization, sera were collected by cardiac puncture after anesthetizing mice, and splenic cells were stained with fluorochrome-labeled antibodies and analyzed with FlowJo software. Anti-SRBC IgG isotypes were measured by ELISA. For anti-SRBC antibody analysis, wells were coated with SRBC membrane protein (20 μg/mL) (Bersee) overnight at 4°C. Wells were blocked with 5% BSA for 1 hour and then incubated with diluted serum for 2 hours at room temperature, followed by incubation with anti-mouse IgG1 (1:3000) (Bethyl Laboratories), IgG2a (1:5000) (Bethyl Laboratories), IgG2b (1:5000) (Bethyl Laboratories), IgG3 (1:3000) (Bethyl Laboratories), and anti-mouse IgM (1:3000) (SouthernBiotech). For anti-dsDNA IgG detection, the mouse anti-dsDNA IgG ELISA kit (Alpha Diagnostic) was used according to the manufacturer’s protocols. Reagents are listed in Supplemental Table 2.