Recombinant Human IL-35 Fc Chimera Protein, CF Summary
|Human IL-27 beta /EBI3
Accession # Q14213
|(GGGS)4||Human IL-12 alpha /p35
Accession # P29459
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|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.
When Recombinant Human IL-12 R beta 2 Fc Chimera (Catalog # 1959-B2) is immobilized at 5 μg/mL (100 μL/well), Recombinant Human IL-35 Fc Chimera (Catalog # 8608-IL) binds with an ED50of 20-120 ng/mL. Recombinant Human IL-35 from the two competitors have 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 and two 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 185 kDa, respectively. The R&D Systems Protein offers a better purity than the competition.
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).
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- Niedbala, W. et al. (2007) Eur. J. Immunol. 37:3021.
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- Liu, Y. et al. (2014) Leukemia. [Epub ahead of print; PMID: 25363669].
- Zhang, X. et al. (2014) Ann. Hematol. [Epub ahead of print; PMID: 25512184].
- Long, J. et al. (2013) Biochem. Biophys. Res. Commun. 430:364.
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