Multiplex Assay Kit for Interleukin 35 (IL35) ,etc. by FLIA (Flow Luminescence Immunoassay)

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Interleukin 35 (IL35) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Interleukin 35 (IL35) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Interleukin 35 (IL35) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Interleukin 35 (IL35) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-101 90
EDTA plasma(n=5) 78-101 82
heparin plasma(n=5) 87-97 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 35 (IL35) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Interleukin 35 (IL35) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Interleukin 35 (IL35) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 96-105% 89-96% 78-101% 83-99%
EDTA plasma(n=5) 93-102% 83-90% 82-93% 79-95%
heparin plasma(n=5) 97-104% 86-97% 95-105% 93-101%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:IL35) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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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
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
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