Multiplex Assay Kit for Receptor Activator Of Nuclear Factor Kappa B (RANk) ,etc. by FLIA (Flow Luminescence Immunoassay)

CD265; TNFRSF11A; TNFRSF11-A; ODFR; OFE; TRANCER; Osteoclast differentiation factor receptor; Tumor Necrosis Factor Receptor Superfamily,Member 11a,NFKB Activator

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Receptor Activator Of Nuclear Factor Kappa B (RANk) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Receptor Activator Of Nuclear Factor Kappa B (RANk) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Receptor Activator Of Nuclear Factor Kappa B (RANk) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Receptor Activator Of Nuclear Factor Kappa B (RANk) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 101
EDTA plasma(n=5) 79-90 83
heparin plasma(n=5) 85-98 89
sodium citrate plasma(n=5) 88-96 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Receptor Activator Of Nuclear Factor Kappa B (RANk) ,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 Receptor Activator Of Nuclear Factor Kappa B (RANk) ,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 Receptor Activator Of Nuclear Factor Kappa B (RANk) ,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) 78-104% 91-104% 90-104% 96-103%
EDTA plasma(n=5) 93-105% 96-104% 87-94% 80-99%
heparin plasma(n=5) 88-98% 78-102% 85-97% 90-99%
sodium citrate plasma(n=5) 96-105% 79-94% 78-101% 86-102%

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#:RANk) 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
European Journal of Immunology Human polymorphonuclear neutrophils express the receptor activator of NFκB (RANK) and are activated by its ligand RANKL Wiley: source
International Journal of Cardiology Valvular osteoclasts in calcification and aortic valve stenosis severity PubMed: 23452891
Bulletin  of  Experimental  Biology  and  Medicine Receptor Activator of Nuclear Transcription Factor NF-κB(RANK), Its Ligand RANKL, and Natural Inhibitor of RANKL Osteoprotegerin (OPG) in the Blood Serum of Patients with Primary Bone Tumors DOI:110.1007/s10517-017-3832-9
BIOSCIENCE REPORTS IL-38 restrains inflammatory response of collagen-induced arthritis in rats via SIRT1/HIF-1α signaling pathway Pubmed: 32347300
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