Multiplex Assay Kit for Interleukin 33 (IL33) ,etc. by FLIA (Flow Luminescence Immunoassay)

DV27; C9ORF26; IL1F11; NFHEV; Interleukin-1 Family, Member 11; Nuclear factor from high endothelial venules

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Interleukin 33 (IL33) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Interleukin 33 (IL33) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Interleukin 33 (IL33) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Interleukin 33 (IL33) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 87-94 90
EDTA plasma(n=5) 96-105 101
heparin plasma(n=5) 97-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 33 (IL33) ,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 33 (IL33) ,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 33 (IL33) ,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) 94-105% 81-101% 79-91% 78-101%
EDTA plasma(n=5) 82-96% 93-105% 78-101% 94-105%
heparin plasma(n=5) 92-101% 98-105% 91-105% 78-96%

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#:IL33) 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
The Korean Journal of Internal Medicine Interleukin-33, matrix metalloproteinase-9, and tissue ınhibitor of matrix metalloproteinase-1 in myocardial infarction PubMed: PMC3604606
Acta Histochem CXCL13 blockade attenuates lupus nephritis of MRL/lpr mice PubMed: 26456520
Journal of Interferon & Cytokine Research Epithelial Cell-Derived Cytokines Contribute to the Pathophysiology of Eosinophilic Chronic Rhinosinusitis Pubmed:26540312
Journal of Interferon & Cytokine Research The Expression and Regulation of Interleukin-33 in Human Epidermal Keratinocytes: A New Mediator of Atopic Dermatitis and Its Possible Signaling Pathway. pubmed:27348082
Archives of Dermatological Research IL-33 circulating serum levels are increased in patients with non-segmental generalized vitiligo pubmed:27388717
Sci Pharm. Chrysin Protects Rat Kidney from Paracetamol-Induced Oxidative Stress, Inflammation, Apoptosis, and Autophagy: A Multi-Biomarker Approach. pubmed:28134775
Biotechnology & Biotechnological Equipment  Association of IL-33, IL1RL1 gene polymorphisms with serum IL-33 levels and risk of asthma in adults and asthmatic bronchitis in children (Chinese) 10.1080:13102818.2018.1471361
Tropical Journal of Pharmaceutical Research Role of IL-33 and ST2 signaling and inflammatory responses in non-small cell lung cancer 172341
Particle and Fibre Toxicology Group II innate lymphoid cells and microvascular dysfunction from pulmonary titanium dioxide nanoparticle exposure Pubmed: 30413212
Scientific Reports Interleukin-33 serum levels in postmenopausal women with osteoporosis Pubmed: 30846811
Cell Reports Tumor-Derived Lactic Acid Contributes to the Paucity of Intratumoral ILC2s Pubmed: 32101749
Allergy Asthma & Immunology Research Relationship of microbial profile with airway immune response in eosinophilic or neutrophilic inflammation of asthmatics Pubmed: 32141256
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
J Immunol Res Inhibition of NF-κB/IL-33/ST2 Axis Ameliorates Acute Bronchiolitis Induced by Respiratory Syncytial Virus 34395633
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
researchsquare Dexmedetomidine Attenuates Hyperalgesia Induced By Brachial Plexus Root Avulsion By Restoring The GLT-1 Function Via PKA Signaling
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