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.The IL-12 p40 monomer shares amino acid sequence homology with the IL-6 receptor.It has been reported that activated PBMC produce a manyfold excess of IL-12 p40 monomer over the bioactive p70 heterodimer.The IL-12 p40 monomer has been reported to inhibit binding of IL-12 p70 to the IL-12 receptor, but with 20X less effectiveness than the IL-12 p70 homodimer.Recombinant human IL-12 p70 (Cat. No. 554613) is supplied as a frozen liquid comprised of 0.22 μm sterile-filtered aqueous buffered solution, containingbovine serum albumin and glycerol, with no preservatives.Purity is ≥ 95% as determined by non-reducing SDS-PAGE, and an absorbance assay based on the Beers-Lambert Law. The endotoxin level is ≤ 0.1 ng per μg of human IL-12 p70, as measured in a chromogenic LAL assay.
Suggested Companion Products
Purified Rat Anti-Human IL-12 p70 20C2RUO 0.5mgCat No: 555065
Biotin Mouse Anti-Human IL-12 (p40/p70) C8.6RUO 0.5mgCat No: 554660
Human IL-12 (p40) ELISA Set RUO 20Plate(s)Cat No: 555171
Human IL-12 (p70) ELISA Set RUO 20Plate(s)Cat No: 555183
Human IL-12 (p40) ELISA Kit II RUO 2Plate(s)Cat No: 551116
Human IL-12 (p70) ELISA Kit RUO 2Plate(s)Cat No: 559258
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Preparation and Storage
Store product at -80°C prior to use or for long term storage of stock solutions.Rapidly thaw and quick-spin product prior to use.Avoid multiple freeze-thaws of product.This preparation contains no preservatives, thus it should be handled under aseptic conditions.
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/pharmingen/protocols for technical protocols.
Upon initial thawing, recombinant human IL-12 p70 (Cat. No. 554613) should be aliquoted into polypropylene microtubes and frozen at -80°C for future use. Alternatively, the product can be diluted in sterile neutral buffer containing not less than 0.5 - 10 mg/mL carrier protein, such as human or bovine albumin, aliquoted and stored at -80°C.For in vitro biological assay use, carrier-protein concentrations of 0.5 - 1 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 25 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 human IL-12 p70 (Cat. No. 554613) can be useful as a quantitative standard for measuring human IL-12 protein levels using sandwich ELISA with the purified 20C2 antibody (Cat. No.555065) as a capture antibody and biotinylated C8.6 antibody (Cat. No. 554660) as the detection antibody.To obtain linear standard curves, investigators may want to consider using doubling dilutions of recombinant human IL-12 from 2,000 - 15 pg/mL to be included in each ELISA plate.For measuring human IL-12 in serum or plasma, investigators are highly encouraged to use the BD OptEIA™ Human IL-12 ELISA Set (Cat. No. 555171 for human IL-12 p40 and Cat. No. 555183 for human IL-12 p70) or the BD OptEIA™ Human IL-12 ELISA Kit II (Cat. No. 551116 for human IL-12 p40 and Cat. No. 559258 for human IL-12 p70).
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.Anactivity range of 0.2 - 5.0 x 10^8 units/mg, encompassing an
ED50= 20 - 500 pg/mL, has previously been reported using PHA/IL-2 -activated human peripheral blood mononuclear cell (PBMC) for proliferation, with a unit defined as the amount of material needed to stimulate a half-maximal response at cytokine saturation.
