Recombinant Human IL-35 Fc Chimera Protein, CF

When Recombinant Human IL-12 R beta 2 Fc Chimera (Catalog # 1959-B2) isimmobilized at 5 μg/mL (100 μL/well), Recombinant Human IL-35 Fc Chimera(Catalog # 8608-IL) binds with an ED50 of20-120 ng/mL. Recombinant Human IL-35from the two competitorshave much weaker Recombinant Human IL-12 R beta 2 Fc Chimera binding activity.

1 μg/lane of Recombinant Human IL-35 Fc Chimera Protein from R&D Systems andtwo competitors was resolved with SDS-PAGE under reducing (R) and non-reducing(NR) conditions and visualized by silver staining. Recombinant Human IL-35 Fc Chimera Protein(Catalog # 8608-IL) from R&D Systems shows single bands at 85 kDa and 185kDa, respectively. The R&D Systems Protein offers a better purity than thecompetition.

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

Recombinant Human IL-35 Fc Chimera Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. When Recombinant Human IL-12 R beta 2 Fc Chimera (Catalog # 1959-B2) is immobilized at 5 μg/mL (100 μL/well), the concentration of Recombinant Human IL-35 Fc Chimera that produces 50% optimal binding response is approximately 20-120 ng/mL.
• Source: Human embryonic kidney cell, HEK293-derived human IL-35 protein
  

  Human IL-27 beta /EBI3 (Arg21-Lys229) Accession # Q14213	(GGGS)4	Human IL-12 alpha /p35 Arg23-Ser219 Accession # P29459	IEGRMD	Human IgG1 Pro100-Lys330)

• Accession #: Q14213 (IL-27 beta /EBI3) & P29459 (IL-12 alpha /p35)
• N-terminal Sequence: Arg21
• Protein/Peptide Type: Recombinant Proteins
• Gene: EBI3
• Purity: >80%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <0.30 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 73 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: 81-97 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.
• Purity: >80%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 100 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-35 Fc Chimera Protein, CF

• IL35
• IL-35

Background

Interleukin 35 (IL-35) is a member of the IL-12 family of heterodimeric cytokines. Unlike other IL-12 family cytokines which stimulate the immune response, the predominant function of IL-35 is as an immunosuppressant. IL-12 cytokines are composed of an alpha and beta subunit which, for IL-35 are the IL-12 p35 subunit and the EBI3 subunit, respectively (1-3). The IL-12 p35 subunit of IL-35 is synthesized as a 219 amino acid (aa) precursor protein with a 22 aa signal sequence and a 197 aa mature region. The EBI3 subunit of IL-35 is synthesized as a 229 aa precursor protein that contains a 20 aa signal sequence and a 209 aa mature region. Human and mouse IL-35 share 58% and 62% sequence homology in their IL-12 p35 and EBI3 subunits, respectively. IL-35 binds to the homodimeric receptors, IL-12 R beta 2 and gp130, as well as to the IL-12 R beta 2-gp130 receptor heterodimer (4). The expression pattern of IL-35 is thought to differ between mouse and humans (5). In mouse regulatory T cells, both subunits of IL-35 are constitutively expressed and the mature IL-35 is secreted. In humans, IL-12 p35 is the only subunit constitutively expressed in regulatory T cells. Immune activation can induce EBI3 expression and IL-35 secretion in human effector T cells (6-8). IL-35 is also expressed and secreted in human placental trophoblasts (1, 9). In both human and mouse IL-35 has been shown to suppress effector T cell proliferation, inhibit Th17 cell development, and promote the conversion of T cells and B cells into regulatory T and B cells, respectively (1, 4, 8, 10, 11). IL-35 is thought to be involved in infectious tolerance and inflammatory cytokine-mediated autoimmune disorders (1, 3, 5, 12). Serum levels of IL-35 are associated with acute graft-versus-host disease following hematopoietic stem cell transplantation (13, 14). IL-35 also functions as a regulator of tumor growth (2, 12, 15).

1. Collison, L.W. and D.A. Vignali (2008) Immunol. Rev. 226:248.
2. Wang, Z. et al. (2013) J. Immunol. 190:2415.
3. Choi, J. et al. (2015) Clin. Rev. Allergy. Immunol.
4. Collison, L.W. et al. (2012) Nat. Immunol. 13:290.
5. Ning-Wei, Z. (2010) Rev. Med. Chil. 138:758.
6. Bardel, E. et al. (2008) J. Immunol. 181:6898.
7. Guttek, K. and D. Reinhold (2013) Cytokine 64:46.
8. Collison, L.W. et al. (2007) Nature 450:566.
9. Mao, H. et al. (2013) Hum. Immunol. 74:872.
10. Wang, R.X. et al. (2014) Nat. Med. 20:633.
11. Niedbala, W. et al. (2007) Eur. J. Immunol. 37:3021.
12. Collison, L.W. et al. (2010) Nat. Immunol. 11:1093.
13. Liu, Y. et al. (2014) Leukemia. [Epub ahead of print; PMID: 25363669].
14. Zhang, X. et al. (2014) Ann. Hematol. [Epub ahead of print; PMID: 25512184].
15. Long, J. et al. (2013) Biochem. Biophys. Res. Commun. 430:364.

Publications for IL-35 (8608-IL)(3)

Publications using 8608-IL

• Feng, J;Li, K;Xie, F;Han, L;Wu, Y; IL-35 ameliorates lipopolysaccharide-induced endothelial dysfunction by inhibiting endothelial-to-mesenchymal transition International immunopharmacology 2024-02-08 [PMID: 38335651] (Bioassay, Human)
• L Lombardell, F Logiodice, O Kullolli, H Haller, C Agostinis, R Bulla, D Rukavina, MP Piccinni At Embryo Implantation Site IL-35 Secreted by Trophoblast, Polarizing T Cells towards IL-35+ IL-10+ IL-4+ Th2-Type Cells, Could Favour Fetal Allograft Tolerance and Pregnancy Success International Journal of Molecular Sciences, 2022-04-28;23(9):. 2022-04-28 [PMID: 35563316] (Bioassay, Human)
• R Xu, RK Shears, R Sharma, M Krishna, C Webb, R Ali, X Wei, A Kadioglu, Q Zhang IL-35 is critical in suppressing superantigenic Staphylococcus aureus-driven inflammatory Th17 responses in human nasopharynx-associated lymphoid tissue Mucosal Immunol, 2020-01-02;13(3):460-470. 2020-01-02 [PMID: 31896761] (Cell Culture, Human)

Bioinformatics

• Gene Symbol: EBI3
• Uniprot:
  • Q14213 (IL-27 beta /EBI3) & P29459 (IL-12 alpha /p35)()