Recombinant Human IL-12 p35 (C96S) Protein, CF

Measured by its ability to induce IFN-gamma secretion in NK-92 human natural killer lymphoma cells, the concentration of Recombinant Human IL‑12 p35 (C96S) Monomer (Catalog # 11209-IL) that produces 50% of the response is 0.300-3.00 ng/mL when Recombinant Human IL‑12 p40 Monomer (309-IL) is present at 0.25 µg/mL.

2 μg/lane of Recombinant Human IL‑12 p35 (C96S) Protein (Catalog # 11209-IL) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 28‑37 kDa.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human IL-12 p35 (C96S) Protein, CF Summary

• Details of Functionality: Measured by its ability to induce IFN-gamma secretion in NK-92 human natural killer lymphoma cells, the concentration of Recombinant Human IL‑12p35 (C96S) Monomer (Catalog # 11209-IL) that produces 50% of the response is 0.300-3.00 ng/mL when Recombinant Human IL‑12p40 Monomer (Catalog # 309-IL) is present at 0.25 µg/mL.
• Source: Human embryonic kidney cell, HEK293-derived human IL-12 protein
  Arg23-Ser219 (Cys96Ser)
• Accession #: P29459.2
• N-terminal Sequence: Arg23
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 23 kDa .
  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: 28-37 kDa, under 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 Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 250 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Human IL-12 p35 (C96S) Protein, CF

• 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, also known as natural killer cell stimulatory factor (NKSF) or cytotoxic lymphocyte maturation factor (CLMF), is a pleiotropic cytokine originally identified in the medium of activated human B lymphoblastoid cell lines (1). The p40 subunit of IL-12 has been shown to have extensive amino acid sequence homology to the extracellular domain of the human IL-6 receptor while the p35 subunit shows distant but significant sequence similarity to IL-6, G-CSF, and chicken MGF (2, 3). These observations have led to the suggestion that IL-12 might have evolved from a cytokine/soluble receptor complex. Human and murine IL-12 share 70% and 60% amino acid sequence homology in their p40 and p35 subunits, respectively. In addition to be a subunit of the proinflammatory cytokine IL-12, p35 is also a subunit of IL-35, an immune suppressor (4). p35 subunit is broadly expressed while p40 subunit is limited to hematopoietic cells (5). Secretion of p35 is dependent on p40 to ensure correct folding (6). Recent studies show that p40 and p35 which are expressed from two different cell lines can assemble to form functional IL-12 in vitro (7). When functions alone, p35 subunit induces expansion of IL-10 and IL-35 expressing regulatory B cells and inhibits neuroinflammation and ameliorates autoimmune encephalomyelitis (8-9).

1. Gubler, U. et al. (1991) Proc. Natl. Acad. Sci. 88:4143.
2. Gearing, D. et al. (1991) Cell 66:9.
3. Merberg, D. et al. (1992) Immunology Today 13:78.
4. Devergne, O. et al. (1997) Proc Natl Acad Sci U.S.A. 94:1204.
5. Carra, G. et al. (2000) J. Immunol 164:4752.
6. Reitberger, S. et al. (2017) J. Biol. Chem 292:8073.
7. Gerber, A.N. et al. (2021) Cell Rep. 37:109816.
8. Dambuza, I.M. et al. (2017) Nat Commun. 8:719.
9. Choi, J.K. et al. (2017) Front Immunol. 8:1258.

Bioinformatics

• Uniprot:
  • P29459.2()