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Human IL-12 P40 monomer Ile23-Ser328 (Accession # NM_002187) was expressed in insect cells.
The 306 amino acid recombinant protein has a predicted molecular mass of 34696.9 Da. The DTT-reduced and the non-reduced protein migrate at approximately 40 kD by SDS-PAGE. The N-terminal amino acid is Isoleucine.
>95%, as determined by Coomassie stained SDS-PAGE.
0.22 µm filtered protein solution is in PBS
Less than 0.01 ng per µg cytokine as determined by the LAL method.
Bottled at 0.1 mg/ml.
Storage & Handling:
Unopened vial can be stored from -20°C to -70°C for 6 months. For maximum results, quick spin vial prior to opening. Stock solutions should be prepared at no less than 1 µg/mL in sterile buffer containing carrier protein such as 1% BSA or HSA or 10% FBS. After dilution, the cytokine can be stored at 4°C for one month or from -20°C to -70°C for up to 3 months. Avoid repeated freeze/thaw cycles.
Each lot of this protein is quality control tested by ELISA assay. For use as an ELISA standard, a standard curve comprised of doubling dilutions from 4000 pg/ml to 30 pg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.
This IL-12 protein is useful as a standard for a human IL-12 sandwich ELISA, using unlabeled C8.3 antibody (catalog #501702) for capture and biotinylated C8.6 antibody (catalog #508802) for detection.
IL-12 and IL-23 share the p40 subunit which heterodimerizes with IL-12 p35 or IL-23 p19 subunits to form IL-12 or IL-23. IL-12 p40 exists as a monomer and as a homodimer (IL-12 p80). IL-12 induction is relevant in asthmatic airway inflammation. Amplified epithelial IL-12 p40 expression and augmented BAL fluid IL-12 p40 (but not IL-12 p70) concentrations has been detected in asthmatic subjects. The IL-12(p40) subunit specifically inhibits IL-12 dependent cellular responses. The IL-12- and IL-23-induced antitumour effects in vivo are abrogated by local production of p40. Genetic polymorphisms in interleukin-12B gene has been associated with systemic lupus erythematosus.
The expression level of p40 is much higher than that of p35 in IL-12p70-producing cells; therefore, monocytes, macrophages, neutrophils, dendritic cells, and B cells express p40 monomer.
p40 monomer and heterodimer are capable of activating microglia and macrophages to induce the production of iNOS and TNF-α. In addition, p40 inhibited IL-23-mediated IFNg and IL-17A production in freshly isolated and Con A-activated splenocytes.
Macrophages, microglia, and PBL.
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