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Human Interleukin-12 Recombinant

IL-12

accession:  P29460
   Size A     10ug   $ 160
   Size B      100ug  $1200
   Size C      500ug  $ 3500
Domain  :  Fibronectin type-III
Gene  :  IL12B
Catalog no. :  RKP29460
Source:
Optimized DNA sequence encoding Human Interleukin-12 mature chain was expressed in HEK cells.

 
Molecular weight:
Native human Interleukin-12, generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 60kDa.

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 39kDa and 43kDA protein under  reducing conditions in SDS-PAGE.


 
Purity:
>97%, as determined by SDS-PAGE and HPLC

 
Biological Activity:
The ED(50) 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.

 
Protein Sequence:
        10         20         30         40         50         60 
MCHQQLVISW FSLVFLASPL VAIWELKKDV YVVELDWYPD APGEMVVLTC DTPEEDGITW 

        70         80         90        100        110        120 
TLDQSSEVLG SGKTLTIQVK EFGDAGQYTC HKGGEVLSHS LLLLHKKEDG IWSTDILKDQ 

       130        140        150        160        170        180 
KEPKNKTFLR CEAKNYSGRF TCWWLTTIST DLTFSVKSSR GSSDPQGVTC GAATLSAERV 

       190        200        210        220        230        240 
RGDNKEYEYS VECQEDSACP AAEESLPIEV MVDAVHKLKY ENYTSSFFIR DIIKPDPPKN 

       250        260        270        280        290        300 
LQLKPLKNSR QVEVSWEYPD TWSTPHSYFS LTFCVQVQGK SKREKKDRVF TDKTSATVIC 

       310        320        330        340        350        360 
RKNASISVRA QDRYYSSSWS EWASVPCS     RNLPVATP DPGMFPCLHH SQNLLRAVSN
       370        380        390        400        410        420 
MLQKARQTLE FYPCTSEEID HEDITKDKTS TVEACLPLEL TKNESCLNSR ETSFITNGSC

       430        440        450        460        470        480 
LASRKTSFMM ALCLSSIYED LKMYQVEFKT MNAKLLMDPK RQIFLDQNML AVIDELMQAL

       490        500        510        520        530        
NFNSETVPQK SSLEEPDFYK TKIKLCILLH AFRIRAVTID RVMSYLNAS
(*)Complete precursor sequence shown, expressed chain highlighted
 
Endotoxin:
Endotoxin content was assayed using a LAL gel clot method.
Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).

 
Presentation:
Recombinant Interleukin-12 was lyophilized from a 0.2 μm filtered PBS solution pH 7.5.

 
Reconstitution:
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.

 
Storage:
The lyophilized protein is stable for at least 2 years from date of receipt at -20° C.
Upon reconstitution, this cytokine can be stored in working aliquots at 2° - 8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity.

Avoid repeated freeze/thaw cycles.

 
Usage:
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.

 

Human IL-12

IL12 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 IL12 are bacteria, bacterial products, and parasites. IL12 is produced after stimulation with Phorbol esters or Calcium ionophore by human B- lymphoblastoid cells. The IL12 receptor ,CD212, appears to be a single protein of approximately 110 kDa. Up to 1000-9000 high affinity IL12 receptors/cell are expressed on peripheral blood mononuclear cells activated by various mitogens for T-cells or by IL2. IL12 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 + IL2, or SAC + IL2 do not express the receptor. Human IL12 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 IL12 is active on both murine and human lymphocytes.

Related Publications:
gc1q receptor ligation selectively down-regulates Human IL-12 production through activation of the phosphoinositide 3-kinase pathway
J. Immunol., Oct 2005; 175: 4706 - 4714.
Human IL-12(p35) gene activation involves selective remodeling of a single nucleosome within a region of the promoter containing critical sp1-binding sites
Blood, Jun 2003; 101: 4894 - 4902.
impaired accumulation and function of memory cd4 t cells in Human IL-12 receptor 1 deficiency
J. Immunol., Jan 2003; 170: 597 - 603.
biosynthesis and posttranslational regulation of Human IL-12
J. Immunol., May 2000; 164: 4752 - 4761.
disparate intracellular processing of Human IL-12 preprotein subunits: atypical processing of the p35 signal peptide
J. Immunol., Jan 2000; 164: 839 - 847.




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