Description
Interleukin-12 (IL-12) is a potent regulator of cell-mediated immune responses. Biologically active IL-12 is secreted by activated B lymphocytes and macrophages as a 70 kD heterodimeric glycoprotein comprised of disulfide-bonded 35 kD (p35) and 40 kD (p40) subunits. IL-12 binds to specific high affinity receptors and can; i) stimulate the growth of activated CD4+ and CD8+ T cells and natural killer (NK) cells; ii) promote the development of proinflammatory/Th1-like CD4+ cells and cytotoxic CD8+ T cells; iii) increase the lytic activity of NK cells, lymphokine-activated killer cells and antibody dependent cell-mediated cytotoxic cells, and; iv) induce NK and T cells to secrete large amounts of the proinflammatory cytokine, IFN-γ. Recombinant mouse IL-12 p70 (Cat. No. 554592) is provided lyophilized, containing bovine serum albumin and no preservatives. Recombinant mouse IL-12 p70 was found to be ≥ 95% pure as determined by SDS-PAGE, and an absorbance assay based on the Beer-Lambert law. The endotoxin level is ≤ 0.1 ng per μg of mouse IL-12 p70, as measured in a chromogenic LAL assay.
Preparation And Storage
Recommended Assay Procedures
Store lyophilized or reconstituted recombinant mouse IL-12 p70 at -80°C. The material may be reconstituted with sterile neutral buffer containing no less than 1 - 10 mg/mL carrier protein, such as human or bovine albumin. Reconstituted mouse IL-12 p70 should be aliquoted and stored at -80°C. For in vitro biological assay use, carrier-protein concentrations of 1 - 2 mg/mL are recommended. For use as an ELISA standard, carrier-protein concentrations of 5 - 10 mg/mL are recommended. Failure to add carrier protein or store at indicated temperatures may result in a loss of activity. The product should not be diluted to less than 50 μg/mL for long term storage. Carrier proteins should be pre-screened for possible effects in each investigator's experimental system. Carrier proteins may have an undesired influence on experimental results due to toxicity, high endotoxin levels or possible blocking activity.
ELISA Standard: Recombinant mouse IL-12 p70 (Cat. No 554592) is useful as a quantitative standard for measuring mouse IL-12 p70 protein levels using sandwich ELISA with purified 9A5 antibody (Cat. No. 554658) as a capture antibody and biotinylated C17.8 antibody (Cat. No. 554476) as the detection antibody. To obtain linear standard curves, doubling dilutions of mouse IL-12 p70 standard from 4000 - 30 pg/mL should be included in each ELISA plate. For measuring mouse IL-12 p70 in serum or plasma, investigators are highly encouraged to use the BD OptEIA™ Mouse IL-12 p70 ELISA Set (Cat. No. 555256).
Bioassay: Investigators are advised that the Bioassay application is not routinely tested for this material and are highly encouraged to both titrate this material and include appropriate controls in relevant experiments. An activity range of 0.2 - 2.0 x 10^9 units/mg, encompassing an
ED50 = 5 - 50 pg/mL has previously been reported using 2D6 as indicator cells for proliferation, with a unit defined as the amount of material needed to stimulate a half-maximal response at cytokine saturation.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Development References (3)
-
Maruo S, Ahn HJ, Yu WG, et al. Establishment of an IL-12-responsive T cell clone: its characterization and utilization in the quantitation of IL-12 activity. J Leukoc Biol. 1997; 61(3):346-352. (Biology). View Reference
-
Schoenhaut DS, Chua AO, Wolitzky AG, et al. Cloning and expression of murine IL-12. J Immunol. 1992; 148(11):3433-3440. (Biology). View Reference
-
Zeh HJ III, Tahara H, Lotze MT. Interleukin-12. In: Thomson AW, ed. The Cytokine Handbook. London: Academic Press Limited; 1994:239-256.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
