Multiplex Assay Kit for Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) ,etc. by FLIA (Flow Luminescence Immunoassay)

CD253; TNFSF10; APO2L; Apo2-L; TL2; TRAIL; Tumor Necrosis Factor Ligand Superfamily Member 10; Apo-2 Ligand

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-101 95
EDTA plasma(n=5) 91-101 96
heparin plasma(n=5) 94-102 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) ,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 Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) ,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 Tumor Necrosis Factor Related Apoptosis Inducing Ligand (TRAIL) ,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) 87-95% 87-103% 93-103% 92-105%
EDTA plasma(n=5) 98-105% 87-101% 85-93% 85-101%
heparin plasma(n=5) 89-103% 79-102% 88-105% 80-96%

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#:TRAIL) 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
PLOS ONE Intranasal Administration of Recombinant TRAIL Down-Regulates CXCL-1/KC in an Ovalbumin-Induced Airway Inflammation Murine Model Pubmed:25506835
PLOS ONE Soluble TRAIL Concentration in Serum Is Elevated in People with Hypercholesterolemia PubMed: 26633016
Scientific Reports Non-alcoholic fatty liver disease, vascular inflammation and insulin resistance are exacerbated by TRAIL deletion in mice pubmed:28507343
Journal of Molecular Medicine TRAIL and TRAIL Receptors as Prognostic Markers in Breast Cancer Patients Pubmed: 31183506
life sciences Nifuroxazide attenuates experimentally-induced hepatic encephalopathy and the associated hyperammonemia and cJNk/caspase-8/TRAIL activation in rats Pubmed: 32259601
Int Immunopharmacol Cardio-protective impact of gabapentin against doxorubicin-induced myocardial toxicity in rats; emphasis on modulation of inflammatory-apoptotic signaling Pubmed: 33199237
ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY Chemo-preventive effect of crocin against experimentally-induced hepatocarcinogenesis via regulation of apoptotic and Nrf2 signaling pathways Pubmed: 32942000
J Biochem Mol Toxicol Tranilast abrogates cisplatin©\induced testicular and epididymal injuries: An insight into its modulatory impact on apoptosis/proliferation 34047436
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