ELISA Kit for Interleukin 35 (IL35) Homo sapiens (Human) Sandwich ELISA

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  • ELISA Kit for Interleukin 35 (IL35) Packages (Simulation)
  • ELISA Kit for Interleukin 35 (IL35) Packages (Simulation)
  • ELISA Kit for Interleukin 35 (IL35) Results demonstration
  • SEC008Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered


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

Matrix Recovery range (%) Average(%)
serum(n=5) 95-105 98
EDTA plasma(n=5) 84-98 93
heparin plasma(n=5) 86-102 90


Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 35 (IL35) 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) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%


The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Interleukin 35 (IL35) 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) 98-105% 82-101% 87-105% 91-98%
EDTA plasma(n=5) 90-101% 91-105% 80-96% 80-105%
heparin plasma(n=5) 78-97% 94-101% 78-97% 87-102%


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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

ELISA Kit for Interleukin 35 (IL35)

Test principle

The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to IL12A. Standards or samples are then added to the appropriate microplate wells with a biotin-conjugated monoclonal antibody specific to IL27B. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain IL35, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of IL35 in the samples is then determined by comparing the O.D. of the samples to the standard curve.


  • Aberrant expression of Treg-associated cytokine IL-35 along with IL-10 and TGF-β in acute myeloid leukemia.PubMed: 22783403
  • Plasma IL-17, IL-35, interferon-γ, SOCS3 and TGF-β levels in pregnant women with preeclampsia, and their relation with severity of diseasePubmed: 24175856
  • What is the impact of Th1/Th2 ratio, SOCS3, IL17, and IL35 levels in unexplained infertility?Pubmed: 24368037
  • The imbalanced profile and clinical significance of T helper associated cytokines in bone marrow microenvironment of the patients with acute myeloid leukemiaScienceDirect: S0198885913005776
  • 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 MothersPubmed:24785110
  • Function of interleukin?17 and ?35 in the blood of patients with hepatitis B?related liver cirrhosisPubmed:25323532
  • The regular distribution and expression pattern of immunosuppressive cytokine IL-35 in mouse uterus during early pregnancyPubmed:25611266
  • Wogonin Inhibits Tumor-derived Regulatory Molecules by Suppressing STAT3 Signaling to Promote Tumor ImmunityPubMed: 25962106
  • Interleukin 35 Synovial Fluid Levels Are Associated with Disease Activity of Rheumatoid ArthritisPubMed: 26204444
  • Interleukin-35 is upregulated in systemic sclerosis and its serum levels are associated with early diseasePubMed: 26231346
  • Anti-inflammatory effects of interleukin-35 in acquired aplastic anemiaPubMed: 26282938
  • Interleukin-35 Inhibits Endothelial Cell Activation by Suppressing MAPK-AP-1 PathwayPubMed: 26085094
  • Interleukin 35 may contribute to the loss of immunological self‐tolerance in patients with primary immune thrombocytopeniaPubMed: 25640666
  • New insights into immune mechanisms underlying response to Rituximab in patients with membranous nephropathy: A prospective study and a review of the literaturePubmed:26876383
  • Increased plasma concentrations of interleukin 35 in patients with coronary artery diseaseJournal:-19
  • The clinical utility of serum IL-35 in patients with polymyositis and dermatomyositispubmed:27502600
  • Enhanced LPS-induced activation of IL-27 signalling in sarcoidosispubmed:27492538
  • Effects of an intravitreal injection of interleukin-35-expressing plasmid on pro-inflammatory andanti-inflammatory cytokines.pubmed:27460435
  • 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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