IL12 Mouse, Sf9
Sf9, Baculovirus cells.
Interleukin 12 (subunit beta/alpha), IL12b/IL12a, Il-12b/Il-12a, IL-12p40/Il-12p35, Il12p40/Il12p35, p40/p35, Sf9.
Greater than 95.0% as determined by SDS-PAGE.
Description
IL12 Mouse Recombinant produced in a baculovirus expression system is a glycosylated disulfide linked (through cysteines in bold) heterodimer comprised of IL12A (23-335aa, total of 319 aa, MW 35.7kDa) and IL12B (23-215aa, total of 199 aa, MW 22.5kDa), having a total predicted molecular mass of 58.3kDa (Molecular weight on SDS-PAGE will appear higher). IL12A is fused to a 6 amino acid His-Tag at C-terminus and purified by proprietary chromatographic techniques.
Product Specs
Interleukin 12, composed of subunits beta (IL12b) and alpha (IL12a), is a cytokine that plays a crucial role in cell-mediated immunity. It enhances the cytotoxic activity of natural killer (NK) cells and lymphocyte-activated killer (LAK) cells, as well as promotes the production of interferon-gamma (IFN-γ) from resting peripheral blood mononuclear cells (PBMCs). IL12b/IL12a is essential for the differentiation of T helper 1 (Th1) cells from precursor T helper cells. In conjunction with IL23A, it forms interleukin-23 (IL-23), which is implicated in the development of autoimmune inflammation, influencing autoimmune inflammatory diseases and tumorigenesis.
Recombinant mouse IL12, produced using a baculovirus expression system in Sf9 insect cells, is a glycosylated heterodimer linked by disulfide bonds (formed through cysteine residues, indicated in bold). It consists of IL12A (amino acids 23-335, total 319 aa, molecular weight 35.7 kDa) and IL12B (amino acids 23-215, total 199 aa, molecular weight 22.5 kDa), resulting in a predicted total molecular mass of 58.3 kDa. However, it's important to note that the apparent molecular weight on SDS-PAGE might be higher. IL12A is tagged with a 6-amino acid His-tag at its C-terminus. The protein is purified using proprietary chromatographic techniques.
The IL12 protein solution is provided at a concentration of 0.5 mg/ml in phosphate-buffered saline (pH 7.4) containing 10% glycerol.
The purity of IL12 is greater than 95%, as determined by SDS-PAGE analysis.
Biological activity is assessed using an IFN-γ ELISA with NK-92 human natural killer cells. The ED50, representing the concentration at which 50% of the maximal effect is observed, is less than or equal to 10 ng/ml.
Interleukin 12 (subunit beta/alpha), IL12b/IL12a, Il-12b/Il-12a, IL-12p40/Il-12p35, Il12p40/Il12p35, p40/p35, Sf9.
Sf9, Baculovirus cells.
IL12B(p40)
MWELEKDVYV VEVDWTPDAP GETVNLTCDT PEEDDITWTS DQRHGVIGSG KTLTITVKEF LDAGQYTCHK GGETLSHSHL LLHKKENGIW STEILKNFKN KTFLKCEAPN YSGRFTCSWL VQRNMDLKFN IKSSSSSPDS RAVTCGMASL SAEKVTLDQR DYEKYSVSCQ EDVTCPTAEE
TLPIELALEA RQQNKYENYS TSFFIRDIIK PDPPKNLQMK PLKNSQVEVS WEYPDSWSTP HSYFSLKFFV RIQRKKEKMK ETEEGCNQKG AFLVEKTSTE VQCKGGNVCV QAQDRYYNSS CSKWACVPCR VRS
IL12A(p35)
RVIPVSGPAR CLSQSRNLLK TTDDMVKTAR EKLKHYSCTA EDIDHEDITR DQTSTLKTCL PLELHKNESC LATRETSSTT RGSCLPPQKT SLMMTLCLGS IYEDLKMYQT EFQAINAALQ NHNHQQIILD KGMLVAIDEL MQSLNHNGET LRQKPPVGEA DPYRVKMKLC ILLHAFSTRV
VTINRVMGYL SSAHHHHHH.
Product Science Overview
Interleukin 12 (IL-12) is a cytokine that plays a crucial role in the regulation of immune responses. It is a heterodimeric protein composed of two subunits, p35 and p40, which are linked by disulfide bonds. The recombinant form of IL-12, produced in Sf9 cells (a cell line derived from the fall armyworm), is used extensively in research and therapeutic applications due to its ability to modulate immune responses.
IL-12 is a glycosylated heterodimer containing 503 amino acids, with the p40 subunit consisting of 306 amino acids and the p35 subunit consisting of 197 amino acids. The molecular mass of the recombinant IL-12 is approximately 60 kDa . The protein is purified using proprietary chromatographic techniques to ensure high purity and stability.
The Sf9 cell line, derived from the fall armyworm (Spodoptera frugiperda), is commonly used for the production of recombinant proteins. These cells are infected with a baculovirus vector containing the genes encoding the p35 and p40 subunits of IL-12. The recombinant IL-12 produced in Sf9 cells is biologically active and retains the ability to modulate immune responses effectively .
IL-12 is a potent regulator of cell-mediated immune responses. It induces the production of interferon-gamma (IFN-γ) by natural killer (NK) cells and T cells, promoting the differentiation of naive T cells into Th1 cells. This cytokine enhances the cytotoxic activity of NK cells and cytotoxic T lymphocytes, making it a critical component in the immune response against intracellular pathogens and tumors .
Recombinant IL-12 has several applications in research and therapy:
- Cancer Immunotherapy: IL-12 is used to enhance the immune response against tumors by promoting the activation and proliferation of cytotoxic T cells and NK cells.
- Vaccine Adjuvant: IL-12 is used as an adjuvant in vaccines to boost the immune response and improve vaccine efficacy.
- Autoimmune Diseases: IL-12 is studied for its role in autoimmune diseases, where it can modulate the immune response and potentially reduce inflammation .
Lyophilized recombinant IL-12 is stable at room temperature for up to three weeks but should be stored desiccated below -18°C for long-term storage. Upon reconstitution, it should be stored at 4°C for short-term use (2-7 days) and below -18°C for long-term use. It is essential to avoid repeated freeze-thaw cycles to maintain the protein’s stability and activity .
