Multiplex Assay Kit for Neuropilin 1 (NRP1) ,etc. by FLIA (Flow Luminescence Immunoassay)

CD304; BDCA4; VEGF165R; Vascular endothelial cell growth factor 165 receptor

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Neuropilin 1 (NRP1) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Neuropilin 1 (NRP1) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Neuropilin 1 (NRP1) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Neuropilin 1 (NRP1) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-96 89
EDTA plasma(n=5) 95-105 101
heparin plasma(n=5) 92-101 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Neuropilin 1 (NRP1) ,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 Neuropilin 1 (NRP1) ,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 Neuropilin 1 (NRP1) ,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) 80-89% 78-101% 86-98% 92-101%
EDTA plasma(n=5) 91-99% 92-103% 95-102% 79-95%
heparin plasma(n=5) 87-101% 79-101% 79-104% 99-105%

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#:NRP1) 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
APMIS Peritumoral brain edema in angiomatous supratentorial meningiomas: an investigation of the vascular endothelial growth factor A pathway PubMed: 23398358
Disease Markers Circulating Soluble Neuropilin-1 in Patients with Early Cervical Cancer and Cervical Intraepithelial Neoplasia Can Be Used as a Valuable Diagnostic Biomarker Pubmed:25873749
JOMO KENYATTA UNIVERSITY OF AGRICULTURE AND TECHNOLOGY (博士毕业论文) Cytokine-CpG Motif Oligodeoxynucleotide Co-inoculation in the Murine Malaria Model handle:123456789
Molecular Neurobiology Reduced Sympathetic Innervation in Endometriosis is Associated to Semaphorin 3C and 3FExpression. pubmed:27558236
Scand J Clin Lab Invest Serum concentrations of neuropilin-1 in women with endometriosis Pubmed: 32069143
Life Sciences Curcumin and LOXblock-1 ameliorate ischemia-reperfusion induced inflammation and acute kidney injury by suppressing the semaphorin-plexin pathway. Pubmed: 32603817
Scientific Reports SARS-CoV2 Induced Biochemical Mechanisms in Liver Damage and Intestinal Lesions.
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