Mouse anti il12

Immunol 2HB-microtiter plates were coated with human anti-TNFα (R&D Systems), mouse anti-IL12 (BD Biosciences), mouse anti-TNFα (R&D Systems) or mouse anti-IL-10 (R&D Systems) and then blocked with 2% BSA-PBS for 2 hr. After washing, culture supernatants or serially diluted sera were added overnight at 4° C. Plates were washed, incubated with biotin-labeled secondary Ab and developed by adding phosphatase-conjugated streptavidin (AKP) followed by p-nitrophenyl phosphatase (pNPP) substrate (Southern Biotech). Optical density was measured using a SpectraMax M5 microplate Reader and SoftMax Pro Acquisition and Analysis software (Molecular Devices).
IgG anti-dsDNA was detected by coating Immunol 2HB-microtiter plates with 4 μg/ml of CT-DNA (Sigma) in DNA binding solution (Abcam) for 4 hr. The plates were blocked with 2% BSA-PBS and incubated with diluted mouse serum as above. Bound Ab was detected with biotin labeled IgG (Invitrogen) followed by phosphate-conjugated streptavidin and pNPP substrate.