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Expressed in Sf9 cells as secreted protein (p40: Accession# NM_008352, p35: Accession#NM_008351).
Molecular Mass:
The mIL-12 consists of two subunits linked via a disulphide bond: P40 (Accession# P43432: Met23-Ser335) and P35 (Accession# NP_032377: Arg23-Ala215). The total predicted molecular weight is 57.48 kD. The non-reduced protein migrates at approximately 55 kD and the DTT-reduced protein produces two bands at approximately 40 kD and 24 kD by SDS-PAGE.
Purity:
95%, as determined by Coomassie stained SDS-PAGE.
Formulation:
The protein was 0.22 µm filtered in 10 mM NaH2PO4, 150 mM NaCl, pH 7.2.
Endotoxin Level:
Less than 0.01 ng per µg cytokine as determined by the LAL method.
Preparation:
10-100 µg sizes are bottled at 200 µg/mL, 500 µg sizes and larger are bottled at the concentration indicated on the vial.
Storage & Handling:
Unopened vial can be stored at -20°C for six months or at -70°C for one year. For maximum results, quick spin vial prior to opening. Stock solutions should be prepared at no less than 10 µg/mL in buffer containing carrier protein such as 1% BSA or HSA or 10% FBS. For long term-storage, aliquot into polypropylene vials and store in a manual defrost freezer. Avoid repeated freeze/thaw cycles.
Activity:
ED50 =0.10 to 0.20 ng/ml, corresponding to a specific activity of 1.0 - 0.5 x 107 units/mg, as determined by the dose dependent stimulation of IFNγ production by mouse splenocytes.
This product is included in our Activation Bundles. Learn more...
COA:
Mouse splenocytes IFNγ production induced by mouse IL-12.
Description:
IL12 (p70) is a disulfide-linked heterodimer composed of unrelated p40 (glycosylated) and p35 subunits. IL-12 acts as a growth factor for activated human T and NK cells, enhance the lytic activity of human NK cells, and stimulate the production of IFNg, by resting human PBMC. IL-12R is formed by two chains, IL-12Rβ1 and IL-12Rβ2. IL-12Rβ1 is associated with the Janus kinase (Jak) Tyk2 and binds IL-12 p40; IL-12Rβ2 is associated with Jak2 and binds either the heterodimer or the p35 chain. Signaling through the IL-12 receptor complex induces phosphorylation, dimerization, and nuclear translocation of several signal transducer and activator of transcription (STAT) family members (STAT1, 3, 4, 5), but most of the biological responses to IL-12 have been attributed to STAT4.
IL-12 is produced by myeloid cells and DCs in response to microbial stimuli, such as those mediated by bacteria, fungi, viruses, and intracellular parasites. IL-12 drives Th1 differentiation and IFNγ production. IL-12 acts as a bridge between innate resistance and adaptive immunity. IL-12 production by activated APC is suppressed by IL-10. In addition, IL-12 production by macrophages is regulated by TNFα and nitric oxide. TLR-4 and TLR-9 can cooperate to increase selectively IL-12 production by murine dendritic cells.
Ligand Receptor:
IL-12 receptor is a heterodimer containing IL-12Rβ1 and IL-12Rβ2 subunits.
Interaction:
Cells of hematopoietic origin express the IL-12R, including NK cells, activated T-cells and dendritic cells.
Cellular Sources:
IL-12 is produced by monocytes, macrophages, neutrophils, dendritic cells and B cells. In the CNS, astrocytes and microglia are the main sources of IL-12.
Antigen References:
1. Schoenhaut DS, et al. 1992. J. Immunol.. 148:3433. 2. Manetti R, et al. 1994. J. Exp. Med. 179:1273. 3. Ireland D,et al.2005. Viral Immunol. 18:397. 4. Moreno SE, et al. 2006. J. Immunol. 177:3218. 5. Lyakh L, et al. 2008.Immunol. Rev..226:112. 6. Theiner G, et al. 2008. Mol. Immunol. 45:244.
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