Recombinant Canine IL-12 Protein

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Summary
Reactivity CaSpecies Glossary
Applications Bioactivity

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Recombinant Canine 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 0.4-2 ng/mL.
Source
Chinese Hamster Ovary cell line, CHO-derived canine IL-12 protein
Canine IL-12 p40
(Ile23-Ser329)
Accession # Q28268
10-His tag
Canine IL-12 p35
(Arg26-Ser222)
(Asp63Glu, Tyr64Leu, Ile65Tyr, Pro66Ser)
Accession # Q28267
10-His tag
N-terminus C-terminus
Accession #
N-terminal Sequence
Ile23 (p40 subunit) & Arg26 (p35 subunit)
Structure / Form
Disulfide-linked heterodimer
Protein/Peptide Type
Recombinant Proteins
Gene
IL12A
Purity
>95%, 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
36.3 kDa (p40), 23.6 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
42 kDa and 28 kDa, reducing conditions

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
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Reconstitution Instructions
Reconstitute at 100 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 Canine 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 the founding member of the IL-12 family of heterodimeric cytokines, which have important immunological functions. IL-12 is composed of two disulfide-linked subunits of 35 kDa and 40 kDa, respectively. The 35 kDa subunit (p35) is an alpha -helical protein homologous to IL-6 and G-CSF. The 40 kDa subunit (p40) contains one fibronectin type III and one Ig C2-like domain, and has a high degree of structural homology to type I cytokine receptors. Whereas p35 subunit is unique to IL-12, the p40 subunit is also utilized in IL-23. Mature canine p35 is a 197 amino acids (aa) protein that is secreted as a heterodimer linked to p40. It contains two potential N-linked glycosylation sites and shares 91%, 86%, 85%, 56%, and 55% aa sequence identity with feline, human, porcine, rat and mouse p35, respectively. Mature p40 contains 307 aa and can exist in multiple forms, including monomer, homodimer, heterodimer linked to p19 (forming IL-23), and heterodimer linked to p35 (forming IL-12). It is upregulated by substances such as LPS and CpG that activate antigen-presenting cells. Mature canine p40 shows 94%, 85%, 84%, 65%, and 65% aa sequence identity to feline, human, porcine, rat and mouse p40, respectively. Cells known to produce IL-12 include macrophages, dendritic cells, monocytes, Langerhans cells, neutrophils, and keratinocytes. The activities of IL-12 are mediated by the receptor complex composed of two type I transmembrane proteins: a 100 kDa ligand-binding subunit (IL-12 R beta 1) and a 130 kDa signal transducing subunit (IL-12 R beta 2). IL-12 facilitates hematopoietic stem cell proliferation, induces NK cell proliferation, promotes IgG production (in mice), and potentiates the expansion and late activation of Th1 CD4+ T cells (1 - 5).

  1. Buttner, M. et al. (1997) Cytokine 10:241.
  2. Park, A.Y. and P. Scott (2001) Scand. J. Immunol. 53:529.
  3. Trinchieri, G. et al. (2003) Immunity 19:641.
  4. Brombacher, F. et al. (2003) Trends Immunol. 24:207.
  5. Lankford, C.S. and D.M. Frucht (2003) J. Leukoc. Biol. 73:49.

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Bioinformatics

Gene Symbol IL12A
Entrez
Uniprot