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Encompassing Amino Acids: 23-529
Amino Acid Sequence: IWELKKDVYVVELDWYPDAPGEMVVLTCDTPEEDGITWTLDQSSEVLGSGKTLTIQVKEFGDAGQYTCHKGGEVLSHSLLLLHKKEDGIWSTDILKDQKEPKNKTFLRCEAKNYSGRFTCWWLTTISTDLTFSVKSSRGSSDPQGVTCGAATLSAERVRGDNKEYEYSVECQEDSACPAAEESLPIEVMVDAVHKLKYENYTSSFFIRDIIKPDPPKNLQLKPLKNSRQVEVSWEYPDTWSTPHSYFSLTFCVQVQGKSKREKKDRVFTDKTSATVICRKNASISVRAQDRYYSSSWSEWASVPCS and RNLPVATPDPGMFPCLHHSQNLLRAVSNMLQKARQTLEFYPCTSEEIDHEDITKDKTSTVEACLPLELTKNESCLNSRETSFITNGSCLASRKTSFMMALCLSSIYEDLKMYQVEFKTMNAKLLMDPKRQIFLDQNMLAVIDELMQALNFNSETVPQKSSLEEPDFYKTKIKLCILLHAFRIRAVTIDRVMSYLNAS
Background: IL-12 is secreted by peripheral lymphocytes after induction. It is produced mainly by B-cells and to a lesser extent by T-cells. The most powerful inducers of IL-12 are bacteria, bacterial products, and parasites. IL-12 is produced after stimulation with phorbol esters or calcium ionophore by human B-lymphoblastoid cells. The IL-12 receptor, CD212, appears to be a single protein of approximately 110 kDa. Up to 1000-9000 high affinity IL-12 receptors/cell are expressed on peripheral blood mononuclear cells activated by various mitogens for T-cells or by IL2. IL-12 receptors are present on activated T-cells expressing CD4 and CD8 and on activated CD56(+) natural killer cells. Resting peripheral blood mononuclear cells, tonsillar B-cells, or tonsillar B-cells activated by anti-IgM/Dx, anti-IgM/Dx + IL-2, or SAC + IL-2 do not express the receptor. Human IL-12 is not active in murine lymphocytes. Hybrid heterodimers consisting of murine p35 and human p40 subunits retain bioactivity on murine cells, however, the combination of human p35 and murine p40 is completely inactive on murine cells. Murine IL-12 is active on both murine and human lymphocytes.
Biological Activity: The ED50 was determined by the dose-dependent proliferation of activated human T-lymphoblasts. cells was found to be in the range of 0.1-0.2 ng/ml.
Description: Recombinant Interleukin-12 is a disulfide-linked heterpdimer protein consisting of 306 amino acid residue p40 subunit and a 197 amino acid residue p35 subunit, and migrates due to glycosylation as an approximately 75 kDa protein under non-reducing and reducing conditions in SDS-PAGE. Optimized DNA sequence encoding human Interleukin-12 mature chain was expressed in Chinese Hamster Ovary (CHO) cells.
Endotoxin Level: <0.1 ng/µg (1 EU/µg), using the LAL gel clot method.
Format: lyophilized protein
Formulation: Lyophilized from 0.2 µm filtered PBS, pH 7.5.
Genbank: P29460
Purity: ≥97% by SDS-PAGE and HPLC
Reconstitution: Reconstitute at 0.1-1.0 mg/ml in distilled water. This solution can then be diluted into other buffers. To maximize product collection from vial surface, vortex briefly and then spin down to recollect the liquid.
Storage Stability: The lyophilized protein is stable for at least 2 years from date of receipt when stored at -20°C. Upon reconstitution, store in working aliquots at +4°C for up to one month, or at -20°C for up to six months, in the presence of a carrier protein. Avoid repeated freeze/thaw cycles.
Uniprot: P29460
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. J. Immunol., Jan 2003, 170: 597 - 603.
2. J. Immunol., May 2000, 164: 4752 - 4761.
3. J. Immunol., Jan 2000, 164: 839 - 847.
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