Western blot analysis of recombinant human IL-18 protein (hIL-18) at indicated levels using IL-18 (D2F3B) Rabbit mAb.
Western blot analysis of extracts from various cell lines using IL-18 (D2F3B) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 263
IL-18 (D2F3B) Rabbit mAb recognizes endogenous levels of total IL-18 protein. This antibody detects 110 and 250 kDa bands of unknown origin in some cell lines.
Monoclonal antibody is produced by immunizing animals with human IL-18 recombinant protein.
Interleukin-18 (IL-18), originally known as interferon-gamma inducing factor (IGIF), is part of the IL-superfamily of cytokines (1,2). This proinflammatory cytokine is synthesized as an inactive precursor which requires cleavage by Caspase-1 to become an active, mature molecule, which is secreted by monocytes and macrophages (3,4,6). It induces IFNγ production in T-helper-1 (Th1) cells and natural killer (NK) cells, and can regulate innate immunity via both Th1 and Th2 responses (4,5). Elevated IL-18 levels are associated with autoimmune disease, but may be balanced by binding to IL-18 binding protein (IL-18BP), which along with IL-18 neutralizing antibodies, are being examined in clinical trials (6,7).
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