Recombinant Rhesus Macaque IL-12 Protein

R&D Systems | Catalog # 3216-RL

R&D Systems
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Key Product Details

  • R&D Systems CHO-derived Recombinant Rhesus Macaque IL-12 Protein (3216-RL)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

CHO

Structure / Form

Disulfide-linked heterodimer

Applications

Bioactivity
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Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived rhesus macaque IL-12 protein
Rhesus Macaque IL-12 (p40)
(Ile23 - Ser328)
Accession # P48095
6-His tag
Rhesus Macaque IL-12 (p35)
(Arg57 - Ser253)
Accession # NP_001038199
6-His tag
N-terminus C-terminus

Purity

>90%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ile23 (p40) & Arg57 (p35)

Predicted Molecular Mass

35.6 kDa (p40), 23.4 kDa (p35)

SDS-PAGE

40-45 kDa and 36-37 kDa, reducing conditions

Activity

Measured in a cell proliferation assay using PHA-stimulated human T lymphoblasts. Symons, J.A. et al. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 272.
The ED50 for this effect is 0.07-0.3 ng/mL.

Formulation, Preparation, and Storage

Carrier Free
What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

Carrier: 3216-RL
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Carrier Free: 3216-RL/CF
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: IL-12

Interleukin 12 (IL-12) is a 70 - 75 kDa heterodimeric glycoprotein that belongs to the IL-12 family of cytokines (1 - 3). Monkey IL-12 consists of two disulfide-linked subunits which are 35 kDa and 40 kDa in size. They show no meaningful amino acid (aa) similarity (4). The mature 35 kDa subunit (p35; also IL-12A) is an alpha -helix, 197 aa in length, that contains seven cysteines and two potential N-linked glycosylation sites (4). Mature Rhesus monkey p35 is 94%, 98%, 97%, 57%, 81%, and 82% aa identical to human, mangabey, baboon, mouse, equine and porcine mature p35, respectively. The mature 40 kDa subunit (p40; alsoIL-12B) is 306 aa in length and contains one intrachain disulfide bond plus three potential N-linked glycosylation sites. Mature Rhesus p40 is 96%, 97%, 99%, 67%, 85%, and 80% aa identical to human, mangabey, baboon, mouse, equine, and porcine p40, respectively. While p35 is reminescent of a hematopoietin ligand, p40 strongly resembles the N-terminus of a hematopoietin receptor, exhibiting a WSXWS motif, an immunoglobulin-like domain, and one fibronectin type III domain (5). This has led to the suggestion that IL-12 may be a cytokine-receptor analog to the IL-6/soluble IL-6R complex (6 - 8). Notably, while p40 may circulate as a monomer or homodimer, p35 is never found by itself. Cells known to produce IL-12 include macrophages and dendritic cells, monocytes, Langerhans cells, neutrophils, and keratinocytes. Functionally, IL-12 has been shown to both enhance cytotoxic activity and induce interferon-gamma (IFN-gamma ) production in NK cells and T cells. In response to infection, IL-27 is released initially, promoting a Th0 to Th0/1 transition state. IL-12 follows next, generating Th1 effector cells. With IL-18, IL-12 creates Th1 memory cells out of effector cells (1 - 3, 6, 7).

References

  1. Hunter, C.A. (2005) Nat. Rev. Immunol. 5:521.
  2. Langrish, C.L. et al. (2004) Immunol. Rev. 202:96.
  3. Kreymborg, K. et al. (2005) Expert Opin. Ther. Targets 9:1123.
  4. Villinger, F. et al. (1995) J. Immunol. 155:3946.
  5. GenBank Accession # P48095.
  6. Brombacher, F. et al. (2003) Trends Immunol. 24:207.
  7. Trinchieri, G. (2003) Nat. Rev. Immunol. 3:133.
  8. Gearing, D.P. and D. Cosman (1991) Cell 66:9.

Long Name

Interleukin 12

Alternate Names

CLMF1, IL12, IL12A, NKSF1

Entrez Gene IDs

3592 (Human); 16159 (Mouse); 84405 (Rat); 397053 (Porcine); 403977 (Canine); 102115316 (Cynomolgus Monkey); 493741 (Feline); 703205 (Rhesus Macaque)

Gene Symbol

IL12A

Additional IL-12 Products

Product Documents for Recombinant Rhesus Macaque IL-12 Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Rhesus Macaque IL-12 Protein

For research use only

Citations for Recombinant Rhesus Macaque IL-12 Protein

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