Multiplex Assay Kit for Ribonuclease A3 (RNASE3) ,etc. by FLIA (Flow Luminescence Immunoassay)

Rnase-A3; RnaseA3; ECP; RNS3; Ribonuclease,RNase A Family 3; Eosinophil Cationic Protein

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Ribonuclease A3 (RNASE3) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Ribonuclease A3 (RNASE3) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Ribonuclease A3 (RNASE3) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Ribonuclease A3 (RNASE3) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 99-105 102
EDTA plasma(n=5) 81-96 85
heparin plasma(n=5) 92-99 95
sodium citrate plasma(n=5) 95-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Ribonuclease A3 (RNASE3) ,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 Ribonuclease A3 (RNASE3) ,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 Ribonuclease A3 (RNASE3) ,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) 87-102% 81-94% 89-98% 94-102%
EDTA plasma(n=5) 90-98% 80-95% 82-89% 78-103%
heparin plasma(n=5) 86-99% 93-101% 96-103% 89-101%
sodium citrate plasma(n=5) 78-91% 89-96% 91-98% 86-96%

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#:RNASE3) 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
Journal of American Science Assessment of the Role of Serum and Urine Eosinophil Cationic Protein in Diagnosis of Wuchereria bancrofti Infection Jofamerican: source
Molecular Medicine Reports Intercellular adhesion molecule?1 expression in activated eosinophils is associated with mucosal remodeling in nasal polyps Pubmed:25573100
International Journal of Molecular Sciences The Human Host Defense Ribonucleases 1, 3 and 7 Are Elevated in Patients with Sepsis after Major Surgery—A Pilot Study Pubmed:26927088
annals of allergy asthma & immunology Interleukin 16 and CCL17/thymus and activation-regulated chemokine in patients with aspirin-exacerbated respiratory disease. pubmed:27986411
European Journal of Clinical Investigation Dying blood mononuclear cell secretome exerts antimicrobial activity. pubmed:27513763
Analytica Chimica Acta An ultrasensitive electrochemical detection of tryptase using 3D macroporous reduced graphene oxide nanocomposites by one-pot electrochemical synthesis
LARYNGOSCOPE Therapeutic Effects of Intranasal Tofacitinib on Chronic Rhinosinusitis with Nasal Polyps in Mice Pubmed: 33014124
INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY Elevated Serum Level of CD48 in Patients with Intermittent Allergic Rhinitis Pubmed: 32966985
Immunopharmacol Immunotoxicol sCD48 is elevated in non-allergic but not in allergic persistent rhinitis 34477021
Mediators Inflamm Roles Played by the PI3K/Akt/HIF-1α Pathway and IL-17A in the Chinese Subtype of Chronic Sinusitis with Nasal Polyps Pubmed:35075349
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