Anti-IL-18 [SAIC-24B-4]

Flow-cytometry using the anti-IL-18 antibody SAIC-24B-4 Human peripheral blood monocytes (PBMC's) were fixed using 2% PFA, permeabilised using 0.5% Triton and stained with unimmunized rabbit IgG antibody (MOPC-21; isotype control, black line) or the rabbit IgG-chimeric version of SAIC-24B-4 (orb613820, blue line) at a dilution of 1:100 for 1h at RT. An additional population of PBMC's stimulated with PMA and Ionomycin in the presence of Monensin for 4 hours were stained in a similar manner (purple line). After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Western Blot using anti-IL-18 antibody SAIC-24B-4 Untreated human peripheral blood monocyte (PBM; A) and PMA/lonomycin-treated PBM lysates (35 µg protein in RIPA buffer) were resolved on a 15% SDS PAGE gel and blots probed with the rabbit IgG-chimeric version of SAIC-24B-4 (orb613820) at 0.3 µg/ml before detection using an anti-rabbit secondary antibody. Protein was detected by chemiluminescence. The expected running size for IL-18 is 22.3kDa. orb613820 successfully detected IL-18 in untreated human peripheral blood monocyte samples.