Multiplex Assay Kit for Synuclein Alpha (SNCa) ,etc. by FLIA (Flow Luminescence Immunoassay)

SNC-A; aSYN; PD1; PARK1; PARK4; NACP; Non-A Beta Component Of Alzheimer's Disease Amyloid Precursor Protein; Non-A4 component of amyloid precursor

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-91 88
EDTA plasma(n=5) 81-91 86
heparin plasma(n=5) 78-88 83

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Synuclein Alpha (SNCa) ,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 Synuclein Alpha (SNCa) ,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 Synuclein Alpha (SNCa) ,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) 98-105% 92-105% 92-103% 93-105%
EDTA plasma(n=5) 85-102% 79-102% 97-105% 87-96%
heparin plasma(n=5) 80-101% 82-92% 89-97% 85-105%

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#:SNCa) 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 Journal of Clinical Investigation Cerebrospinal fluid–based kinetic biomarkers of axonal transport in monitoring neurodegeneration PubMed: PMC3428100
Neurobiology of Disease Lysosomal dysfunction increases exosome-mediated alpha-synuclein release and transmission PubMed: PMC3107939
Nature Communications Parkinson's disease induced pluripotent stem cells with triplication of the α-synuclein locus PubMed: PMC3265381
Provisional chapter Mutations of PARK Genes and Alpha-Synuclein and Parkin Concentrations in Parkinson’s Disease Intechopen:Source
Nerodegeneration Apolipoprotein Eε4: A Biomarker for Executive Dysfunction among Parkinson's Disease Patients with Mild Cognitive Impairment pubmed:29326545
Experimental gerontology The anti-aging protein klotho alleviates injury of nigrostriatal dopaminergic pathway in 6-hydroxydopamine rat model of Parkinson's disease: Involvement of PKA/CaMKII/CREB signaling. pubmed:29107062
Neurobiology of Disease Development and biochemical characterization of a mouse model of Parkinson's disease bearing defective glucocerebrosidase activity Pubmed: 30521842
molecular and cellular biochemistry Potential therapeutic effects of antagonizing adenosine A2A receptor, curcumin and niacin in rotenone-induced Parkinson's disease mice model Pubmed: 31820278
International Journal of Pharmacology Nose to brain delivery of rotigotine loaded chitosan nanoparticles in human SH-SY5Y neuroblastoma cells and animal model of Parkinson's disease Pubmed: 32084576
EBioMedicine Anti-α-synuclein ASO delivered to monoamine neurons prevents α-synuclein accumulation in a Parkinson's disease-like mouse model and in monkeys Pubmed: 32810825
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