Recombinant Human IL-35 Fc Chimera Protein, CF

Catalog # Availability Size / Price Qty
8608-IL-050
Recombinant Human IL-35 Fc Chimera Protein, CF Bioactivity
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Recombinant Human IL-35 Fc Chimera Protein, CF Summary

Purity
>80%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<0.30 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 # 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)
N-terminal Sequence
Analysis
Arg21
Structure / Form
Disulfide-linked homodimer
Predicted Molecular Mass
73 kDa
SDS-PAGE
81-97 kDa, reducing conditions

Product Datasheets

8608-IL

Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Data Images

Bioactivity Recombinant Human IL-35 Fc Chimera Protein, CF Bioactivity View Larger

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 of
20-120 ng/mL. Recombinant Human IL-35from the two competitorshave much weaker Recombinant Human IL-12 R beta 2 Fc Chimera binding activity.

SDS-PAGE Recombinant Human IL-35 Fc Chimera Protein, CF SDS-PAGE View Larger

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.

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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

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Primary Antibodies

Human IgG1 Antibody

Human IgG1 Antibody

Recombinant Proteins

Reconstitution Buffers

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