• Purity

  >80%, 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.  

• Activity

  Measured by its binding ability in a functional ELISA. When Recombinant Human IL-12 R beta 2 Fc Chimera (Catalog # ) 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-27 beta /EBI3 (Arg21-Lys229) Accession # Q14213	(GGGS)4	Human IL-12 alpha /p35 Arg23-Ser219 Accession # P29459	IEGRMD	Human IgG1 Pro100-Lys330)

• N-terminal Sequence    
  Analysis

  Arg21

• Structure / Form

  Disulfide-linked homodimer

• Predicted Molecular Mass

  73 kDa

• SDS-PAGE

  81-97 kDa, reducing conditions

Data Images

Background: IL-35

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).

• References:

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.

• Long Name:

  Interleukin 35

• Entrez Gene IDs:

  10148 (Human); 50498 (Mouse); 680609 (Rat)

• Alternate Names:

  IL35; IL-35