Active Interleukin 35 (IL35)

ACTIVITY TEST

IL35 (Interleukin 35) is an IL-12 family cytokine, which is a dimeric protein composed of IL-12α and IL-27β chains. IL35 is thought to mediate the immune inhibitory function of regulatory T cells and has been proven to promotes pancreas cancer growth through enhancement of proliferation and inhibition of apoptosis. Thus, proliferation assay of IL35 was conducted using PANC-1 cells. Briefly,
PANC-1 cells were seeded into triplicate wells of 96-well plates at a density of 2,000 cells/well and allowed to attach overnight, then the medium was replaced with serum-free standard DMEM prior to the addition of various concentrations of recombinant human IL35. After incubated for 48h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10µL of CCK-8 solution was added to each well of the plate, then the absorbance at 450nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37°C. Proliferation of PANC-1 cells after incubation with IIL35 for 48h observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with human recombinant IL35 for 48h. The result was shown in Figure 2. It was obvious that IL35 significantly increased cell viability of PANC-1 cells.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Oncology Letters Aberrant expression of Treg-associated cytokine IL-35 along with IL-10 and TGF-β in acute myeloid leukemia. PubMed: 22783403
Journal of Maternal-Fetal and Neonatal Medicine Plasma IL-17, IL-35, interferon-γ, SOCS3 and TGF-β levels in pregnant women with preeclampsia, and their relation with severity of disease Pubmed: 24175856
Journal of Reproductive Immunology What is the impact of Th1/Th2 ratio, SOCS3, IL17, and IL35 levels in unexplained infertility? Pubmed: 24368037
Human Immunology The imbalanced profile and clinical significance of T helper associated cytokines in bone marrow microenvironment of the patients with acute myeloid leukemia ScienceDirect: S0198885913005776
Folia Biologica (Praha) Gene Expression of Subunits of the IL-12 Family Cytokines in moDCs Derived In Vitro from the Cord Blood of Children of Healthy and Allergic Mothers Pubmed:24785110
Molecular Medicine Reports Function of interleukin?17 and ?35 in the blood of patients with hepatitis B?related liver cirrhosis Pubmed:25323532
Rom J Morphol Embryol. The regular distribution and expression pattern of immunosuppressive cytokine IL-35 in mouse uterus during early pregnancy Pubmed:25611266
Journal of Immunotherapy Wogonin Inhibits Tumor-derived Regulatory Molecules by Suppressing STAT3 Signaling to Promote Tumor Immunity PubMed: 25962106
PLOS ONE Interleukin 35 Synovial Fluid Levels Are Associated with Disease Activity of Rheumatoid Arthritis PubMed: 26204444
Rheumatology (Oxford) Interleukin-35 is upregulated in systemic sclerosis and its serum levels are associated with early disease PubMed: 26231346
Cytokine Anti-inflammatory effects of interleukin-35 in acquired aplastic anemia PubMed: 26282938
J Biol Chem Interleukin-35 Inhibits Endothelial Cell Activation by Suppressing MAPK-AP-1 Pathway PubMed: 26085094
Br J Haematol Interleukin 35 may contribute to the loss of immunological self‐tolerance in patients with primary immune thrombocytopenia PubMed: 25640666
Autoimmunity Reviews New insights into immune mechanisms underlying response to Rituximab in patients with membranous nephropathy: A prospective study and a review of the literature Pubmed:26876383
Clinical research Increased plasma concentrations of interleukin 35 in patients with coronary artery disease Journal:-19
Clinical Rheumatology The clinical utility of serum IL-35 in patients with polymyositis and dermatomyositis pubmed:27502600
Respiratory Medicine Enhanced LPS-induced activation of IL-27 signalling in sarcoidosis pubmed:27492538
International Journal of Molecular Medicine Effects of an intravitreal injection of interleukin-35-expressing plasmid on pro-inflammatory andanti-inflammatory cytokines. pubmed:27460435
上海口腔医学 IL-35在牙周炎及口腔扁平苔藓中的表达及相关性分析 QK:98604X
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