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Active Interleukin 35 (IL35)

ACTIVITY TEST

IL-35, also known as Interleukin-35, is a heterodimeric cytokine composed of two subunits, p35 and Epstein-Barr virus-induced gene 3 (EBI3). It belongs to the IL-12 family of cytokines and is primarily associated with the regulation of immune responses. IL-35 is produced by regulatory T cells (Tregs) and has been implicated in the suppression of immune responses, playing a role in maintaining immune tolerance and preventing autoimmune diseases. It is also produced by B regulatory cells and can contribute to the suppression of immune reactions in certain contexts.Besides,Interleukin 23 Subunit Alpha (IL23a) has been identified as an interactor of IL-35, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant mouse IL-35 and recombinant human IL23a. Briefly, IL-35 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to IL23a-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-IL-35 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant mouse IL-35 and recombinant human IL23a was shown in Figure 1, the EC50 for this effect is 0.049ug/mL.

USAGE

Reconstitute in 10mM PBS (pH7.4) 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
Reproductive Sciences Upregulation of Interleukin 35 in Patients With Endometriosis Stimulates Cell Proliferation pubmed:28659009
EXPERIMENTAL AND THERAPEUTIC MEDICINE IL‑35 may maintain homeostasis of the immune microenvironment in periodontitis issn:2542-3975
The Journal of Rheumatology Synoviocytes-derived Interleukin 35 Potentiates B Cell Response in Patients with Osteoarthritis and Rheumatoid Arthritis pubmed:29247146
Cellular & Molecular Immunology Human immunodeficiency virus type-1 induces a regulatory B cell-like phenotype in vitro pubmed:28713164
Molecular Medicine Reports Lower level of IL‑35 and its reduced inhibition in Th17 cells in patients with bone marrow mononuclear cells Coombs test‑positive hemocytopenia. pubmed:29257310
Asian pacific journal of allergy and immunology Evaluation of serum interleukin- 35 level in children with persistent asthma. pubmed:27996283
PLoS One Interleukin-35 mitigates the function of murine transplanted islet cells via regulation of Treg/Th17 ratio pubmed:29236782
Hepatology Communications Significant association between FOXP3 gene polymorphism and steroid-resistant acute rejection in living donor liver transplantation 10.1002/hep4.1052
International Journal of Clinical and Experimental Medicine Clinical significance of IL-35 expression in the progression of clear cell renal cell carcinoma ISSN:1940-5901
OncoTargets and Therapy Interleukin-35 as a predictor of prostate cancer in patients undergoing initial prostate biopsy. pubmed:28761357
Annals of Rheumatic Diseases  AB0734 Serum il-35 levels in systemic sclerosis and relationship with clinical features 10.1136:annrheumdis-2018-eular.2605
Tohoku Journal of Experimental Medicine Interleukin-35 as a New Biomarker of Renal Involvement in Lupus Nephritis Patients Pubmed:29576585
Turkish Journal Of Medical Sciences The impact of plasma SOCS3 levels and endometrial leukocytes on unexplained infertility Pubmed:29914245
Oncotarget Pro-tumoral immune cell alterations in wild type and Shb-deficient mice in response to 4T1 breast carcinomas Pubmed:29721156
Journal of Clinical Rheumatology Serum Interleukin 35 Levels in Systemic Sclerosis and Relationship With Clinical Features. Pubmed: 30431486
Oncology Letters Interleukin‑35 is associated with the tumorigenesis and progression of prostate cancer
Journal of Clinical Investigation Plasmacytoid dendritic cells protect against immune-mediated acute liver injury via IL-35 Pubmed: 31264967
Cells CD4+ FOXP3+ T Cells in Rheumatoid Arthritis Bone Marrow Are Partially Impaired Pubmed: 32111105
Cytokine Interleukin-35 in idiopathic inflammatory myopathies Pubmed: 33128920
FEBS Open Bio IL35 predicts prognosis in gastric cancer and is associated with angiogenesis by altering TIMP1, PAI1, and IGFBP1 Pubmed: 33064893
Cancer Cell International Interleukin-35 promotes progression of prostate cancer and inhibits anti-tumour immunity Pubmed: 33041668
Journal of Neuroinflammation Interleukin-9 regulates macrophage activation in the progressive multiple sclerosis brain Pubmed: 32375811
Int Immunopharmacol Association of IL-35 expression and gene polymorphisms in rheumatoid arthritis 33307515
Archives of Medical Science Decreased interleukin-35 levels and CD4+ EBI3+ T cells in patients with type 1 diabetes and the effects of the antibody against CD20 (rituximab) 33488880
Asia Pac Allergy Anti-inflammatory roles of interleukin-35 in the pathogenesis of Japanese cedar pollinosis 34386410
Int Immunopharmacol Pentosan Polysulfate Sodium augments the therapeutic effect of 5-Aminosalicylic Acid in DSS colitis model; the role of IL-35 expression Pubmed:35247859
Biology and Medical Researches Association of three single nucleotide polymorphisms of interleukin-35 for detection and prediction of women with breast cancer in Erbil city
Research Square Analysis of IL-10 and IL-35 in DPP-4 inhibitor-related bullous pemphigoid
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