The recombinant human IL- 12 (IL-12A & IL-12B) protein is expressed in Human Cells, amino acid sequence derived from p35 subunit of human IL-12, termed as IL12A (NP_000873.2), (Met 1-Ser 219) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 1 ; amino acid sequence derived from p40 subunit of human IL-12, termed as IL12B (NP_002178.2), (Met 1-Ser 328) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the heterodimer was purified.
The recombinant IL12 heterodimer of human IL12A/IL12B comprises 525 (208 + 317) amino acids and has a calculated molecular mass of 60.2 (24 + 36.2) kDa. The apparent molecular mass of rh IL12 heterodimer is approximately 40 & 43 kDa respectively in SDS-PAGE under reducing conditions.
Human IL- 12 (IL-12A & IL-12B) protein measured by its ability to bind biotinylated recombinant human IL12RB1 in a functional ELISA.
Measured in a cell proliferation assay using Anti-CD28 and Anti-CD3 stimulated PBMC. The ED50 for this effect is typically 0.1-0.6 ng/mL.
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