Multiplex Assay Kit for Interleukin 1 Receptor Antagonist (IL1RA) ,etc. by FLIA (Flow Luminescence Immunoassay)

IL1RN; IL1-RA; ICIL1RA; IL1ra3; IL1F3; IRAP;IL-1RA; Interleukin-1 Family Member 3; IL1 inhibitor; Anakinra

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Interleukin 1 Receptor Antagonist (IL1RA) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Interleukin 1 Receptor Antagonist (IL1RA) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Interleukin 1 Receptor Antagonist (IL1RA) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Interleukin 1 Receptor Antagonist (IL1RA) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 99-105 102
EDTA plasma(n=5) 94-102 97
heparin plasma(n=5) 82-103 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Interleukin 1 Receptor Antagonist (IL1RA) ,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 1 Receptor Antagonist (IL1RA) ,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 1 Receptor Antagonist (IL1RA) ,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) 93-104% 91-98% 83-101% 93-101%
EDTA plasma(n=5) 90-99% 80-98% 78-98% 99-105%
heparin plasma(n=5) 96-103% 88-96% 98-105% 95-104%

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#:IL1RA) 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 Veterinary Journal Mucosal imbalance of interleukin-1β and interleukin-1 receptor antagonist in canine inflammatory bowel disease ScienceDirect: S1090023312000962
Daily quercetin supplementation over a period of two weeks results in a moderate accumulation of total plasma flavonols in horses Researchgate:Source
Journal of Equine Veterinary Science Synovial Fluid and Serum Concentrations of Interleukin-1 Receptor Antagonist and Interleukin-1 Science: Article
Journal of Equine Veterinary Science A Two-Week Quercetin Supplementation in Horses Results in Moderate Accumulation of Plasma Flavonol Concentrations Science: Article
Scientific publications in peer reviewed journals Evidence-based review of efficacy and adverse effects of joint medication and evaluation of synovial fluid and serum markers for osteoarthritis in the horse receive:101658
Multiple Sclerosis and Related Disorders Interleukin 1 receptor antagonist and 2′-5′-oligoadenylate synthetase-like molecules as novel biomarkers for multiple sclerosis patients in Bahrain pubmed:29141788
Advances in Medical Sciences A similar pro/anti-inflammatory cytokine balance is present in the airways of competitive athletes and non-exercising asthmatics pubmed:28822267
Archives of Medical Research Winter ambient training conditions are associated with increased bronchial hyperreactivity and with shifts in serum innate immunity proteins in young … Pubmed:29379533
Journal of Veterinary Internal Medicine Effect of interleukin‐1β on occludin mRNA expression in the duodenal and colonic mucosa of dogs with inflammatory bowel disease Pubmed:29572935
Frontiers in Pharmacology M10, a Myricetin-3-ObD-Lactose Sodium Salt, Prevents Ulcerative Colitis Through Inhibiting Necroptosis in Mice Pubmed: 33041798
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