Recombinant Rhesus Macaque IL-12 Protein

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Summary
Reactivity RMSpecies Glossary
Applications Bioactivity

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Recombinant Rhesus Macaque IL-12 Protein Summary

Details of Functionality
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.
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
Accession #
N-terminal Sequence
Ile23 (p40) & Arg57 (p35)
Structure / Form
Disulfide-linked heterodimer
Protein/Peptide Type
Recombinant Proteins
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
35.6 kDa (p40), 23.4 kDa (p35).
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
40-45 kDa and 36-37 kDa, reducing conditions
Publications
Read Publications using
3216-RL in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Purity
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 10 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Rhesus Macaque IL-12 Protein

  • CLMF p35
  • CLMF1
  • Cytotoxic lymphocyte maturation factor 35 kDa subunit
  • IL-12 subunit p35
  • IL12
  • IL-12
  • IL-12, subunit p35
  • IL12A
  • interleukin 12, p35
  • interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocytematuration factor 1, p35)
  • interleukin-12 alpha chain
  • interleukin-12 subunit alpha
  • natural killer cell stimulatory factor 1, 35 kD subunit
  • NF cell stimulatory factor chain 1
  • NK cell stimulatory factor chain 1
  • NKSF1
  • p35

Background

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).

  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.

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Publications for IL-12 (3216-RL)(2)

We have publications tested in 1 confirmed species: Primate - Macaca mulatta (Rhesus Macaque).

We have publications tested in 1 application: Bioassay.


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(2)
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Primate - Macaca mulatta (Rhesus Macaque)
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