Multiplex Assay Kit for Interleukin 25 (IL25) ,etc. by FLIA (Flow Luminescence Immunoassay)

IL17E; IL17-E; Interleukin-17E

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Interleukin 25 (IL25) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Interleukin 25 (IL25) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Interleukin 25 (IL25) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Interleukin 25 (IL25) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-101 86
EDTA plasma(n=5) 98-105 102
heparin plasma(n=5) 80-103 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 25 (IL25) ,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 25 (IL25) ,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 25 (IL25) ,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) 80-104% 81-91% 94-103% 98-105%
EDTA plasma(n=5) 91-101% 79-103% 86-101% 79-97%
heparin plasma(n=5) 97-105% 80-102% 82-97% 97-105%

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#:IL25) 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
Am J Clin Exp Immunol. Molecular analysis of interleukin-25 exons 1 and 2 and its serum levels in Iranian patients with multiple sclerosis Pubmed:Pmc4138132
Journal of Interferon & Cytokine Research Epithelial Cell-Derived Cytokines Contribute to the Pathophysiology of Eosinophilic Chronic Rhinosinusitis Pubmed:26540312
Cellular & Molecular Immunology IL-25 stimulates M2 macrophage polarization and thereby promotes mitochondrial respiratory capacity and lipolysis in adipose tissues against obesity. pubmed:28194019
Allergology International The effect of calprotectin on TSLP and IL-25 production from airway epithelial cells pubmed:27475624
Clinical Immunology A mechanism of interleukin-25 production from airway epithelial cells induced by Japanese cedar pollen Pubmed:29402661
Clinical Immunology Toll-like receptor 9 ligands increase type I interferon induced B-cell activating factor expression in chronic rhinosinusitis with nasal polyposis Pubmed:30056130
cyberleninka КЛИНИКО-ЭПИДЕМИОЛОГИЧЕСКАЯ ХАРАКТЕРИСТИКА И ОСОБЕННОСТИ БАЛЛАНСА ЦИТОКИНОВ ПРИ КЛЕЩЕВОМ ЭНЦЕФАЛИТЕ В …
OncoTargets and Therapy effect of Th9/il-9 on the growth of gastric cancer in nude mice Pubmed: 30988627
Cancer Medicine IL‐25 promotes cisplatin resistance of lung cancer cells by activating NF‐κB signaling pathway to increase of major vault protein Pubmed: 31044552
Traditional Medicine Research Qingfei oral liquid downregulates TRPV1 expression to reduce airway inflammation and mucus hypersecretion injury caused by respiratory syncytial virus infection …
Aggravation of Airway Inflammation in RSV-Infected Asthmatic Mice Following Infection-Induced Alteration of Gut Microbiota
Pathogens Elevated Levels of IL-33, IL-17 and IL-25 Indicate the Progression from Chronicity to Hepatocellular Carcinoma in Hepatitis C Virus Patients Pubmed:35056005
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