AdipoGen Life Sciences

IL-27 (mouse):Fc-KIH (human) (rec.)

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AG-40B-0236-C05050 µgCHF 440.00

Figure 1: Protein structure of IL-27 (mouse):Fc-KIH (human) (rec.) (Prod. No. AG-40B-0236) containing a single heterodimer.

Figure 2: IL-27 (mouse):Fc-KIH (human) (rec.) (#AG-40B-0236) activates phosphorylation of Stat3 in HepG2 cells. Method: 5x10⁴ HepG2 cells (a human hepatocellular carcinoma cell line) are treated with IL-27 (mouse):Fc-KIH (human) (rec.) (#AG-40B-0236) at different concentrations indicated on the figure for 15 minutes. Cells are lysed and cell extracts are separated by SDS-PAGE (12.5%) under reducing conditions, transferred to nitrocellulose overnight and incubated with anti-Phospho-Stat3 (Tyr705) (human), Rabbit Monoclonal (RM261) (#REV-31-1142-00) (1/1000) or anti-Stat3, Rabbit Monoclonal (RM465) (#REV-31-1357-00) (1/1’000). After the addition of an anti-rabbit secondary antibody coupled to HRP (1/10’000), proteins are visualized by a chemiluminescence detection system.

Figure 3: IL-27 (mouse):Fc-KIH (human) (rec.) (#AG-40B-0236) (Heterodimer) is more active than an IL-27 (mouse):Fc from a competitor (Dimer of Heterodimer). Method: 5x10⁴ HepG2 cells (a human hepatocellular carcinoma cell line) are treated with IL-27 (mouse):Fc-KIH (human) (rec.) (#AG-40B-0236, AdipoGen) or from IL-27 (mouse):Fc (human) from a competitor at different concentrations indicated on the figure for 15 minutes. Cells are lysed and cell extracts are separated by SDS-PAGE (12.5%) under reducing conditions, transferred to nitrocellulose overnight and incubated with anti-Phospho-Stat3 (Tyr705) (human), Rabbit Monoclonal (RM261) (#REV-31-1142-00) (1/1000) or anti-Stat3, Rabbit Monoclonal (RM465) (#REV-31-1357-00) (1/1’000). After the addition of an anti-rabbit secondary antibody coupled to HRP (1/10’000), proteins are visualized by a chemiluminescence detection system. A) Western Blot, B) Quantification figure of the Western Blot.

Specifications / Handling

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Product Details
Synonyms	InVivoKine™; IL-27 (mouse):Fc Knobs-into-Holes (human) (rec.); Interleukin-27 Subunit α; IL-27 Subunit α; IL-27-A; Interleukin-27 Subunit β; Ebi3; Epstein-Barr Virus-induced Gene 3 Protein Homolog
Product Type	Protein
Properties
Source/Host	CHO cells. Produced using animal component-free medium.
Sequence	IL-27A/p28 (aa 29-234) (mouse):Fc Knobs and IL-27B/EBI3 (aa 19-228) (mouse):Fc Holes form the IL-27 (mouse):Fc-KIH (human) using the Knobs-into-Holes technology (see reference: J.B. Ridgway, et al.; Protein Eng. 9, 617 (1996)).
Crossreactivity	Human Mouse
Specificity	Binds to the mouse and human Il-27 receptor complex.
Biological Activity	Activates STAT3 phosphorylation in mouse and human cells.
MW	~62kDa and 60 kDa (SDS-PAGE)
Purity	≥95% (SDS-PAGE)
Endotoxin Content	<0.01EU/μg purified protein (LAL test).
Concentration	1mg/ml after reconstitution.
Reconstitution	Reconstitute with 50μl sterile water.
Formulation	Lyophilized. Contains PBS
Protein Negative Control	Fc-KIH (human) IgG1 Control (rec.) (AG-35B-0015)
Other Product Data	UniProt link Q8K3I6: IL-27A (mouse) UniProt link O35228 : IL-27B (mouse)
Shipping and Handling
Shipping	BLUE ICE
Short Term Storage	+4°C
Long Term Storage	-20°C
Handling Advice	After reconstitution, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles. Centrifuge lyophilized vial before opening and reconstitution. PBS containing at least 0.1% BSA should be used for further dilutions.
Use/Stability	Stable for at least 6 months after receipt when stored at -20°C. Working aliquots are stable for up to 3 months when stored at -20°C.
Documents
MSDS	Download PDF
Product Specification Sheet
Datasheet	Download PDF

Product-specific References

Description

Interleukin-12 (IL-12) family members are heterodimer glycoproteins, composed of two covalently linked subunits, α and β chains. The α-subunit consists of IL-23p19, IL-27p28, and IL-12p35, and the β-subunit includes IL-12p40 and Epstein-Barr virus-induced gene (Ebi3). IL-12 members bind to cognate heterodimeric receptor chains expressed on T cells.

This family includes IL-12, IL-23, IL-27 and IL-35 and IL-39. IL-12 and IL-23 are predominantly proinflammatory cytokines that contribute key roles in the development of Th1 and Th17 cells, respectively. IL-27 has both pro- and anti-inflammatory properties and is a potent T cell immunomodulator. IL-35, a new member of this family, is a potent inhibitory cytokine produced by natural, thymus-derived regulatory T cell (nTreg) populations. IL-39, the newest member of IL-12 family, mediates the inflammatory response through the activation of STAT1/STAT3 signaling pathway. These IL-12 family members link innate immunity with the development of adaptive immunity and are also important for regulating T cell responses.

IL-27 is composed of two subunits, IL-27p28 and Epstein-Barr virus-induced gene 3 (EBI3) signals via a heterodimeric receptor consisting of WSX-1 and glycoprotein (gp130), leading to activation of STAT 1 and 3. IL-27 is pro-inflammatory and promotes NK and T cell proliferation as well as the production of IFN-γ. It is anti-inflammatory, inhibiting Th2 and Th17 cell activities and stimulating the production of IL-10 by T regulatory cells. IL-27 has potent antiviral activities against numerous viruses, by increasing the production of interferons (IFNs). Finally, IL-27 has antitumor activity as well as anti-angiogenic activity with activation of the production of anti-angiogenic chemokines.

The protein IL-27 (mouse):Fc-KIH (human) (rec.) is produced by using two different vectors, one encoding for the IL-27A/p28:Fc Knobs sequence (synthesizing a protein of 62kDa) and one encoding for the IL27B/EBI3: Fc Holes sequence (synthesizing a protein of 60kDa). Both vectors transfected into CHO cells produce both Fc molecules (Knobs-into-Holes technology; J.B. Ridgway, et al.; Protein Eng. 9, 617 (1996)) required for dimerization and for secretion of the final protein IL-27 (mouse): Fc-KIH (human) (rec.).

InVivoKines™ are a new generation of recombinant fusion proteins for immunotherapeutic, preclinical and translational in vivo research, developed and manufactured by AdipoGen Life Sciences.

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