Multiplex Assay Kit for Acetylcholinesterase (ACHE) ,etc. by FLIA (Flow Luminescence Immunoassay)

ARAChE; N-AChE; YT

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 81-97 89
EDTA plasma(n=5) 78-99 91
heparin plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Acetylcholinesterase (ACHE) ,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 Acetylcholinesterase (ACHE) ,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 Acetylcholinesterase (ACHE) ,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) 85-104% 95-105% 88-97% 88-101%
EDTA plasma(n=5) 79-97% 88-101% 86-94% 90-102%
heparin plasma(n=5) 99-105% 92-101% 89-103% 86-98%

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#:ACHE) 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
Acta Histochem Cytochem. The Effects of Exercise-induced Fatigue on Acetylcholinesterase Expression and Activity at Rat Neuromuscular Junctions PubMed: PMC2775104
Applied Biological Chemistry Positive enhancement of Lactobacillus fermentum HY01 on intestinal movements of mice having constipation 10.1007/s13765-017-0327-3
Molecular Nutrition & Food Research Thymoquinone Can Improve Neuronal Survival and Promote Neurogenesis in Rat Hippocampal Neurons Pubmed:29277983
Inflammation Feishu Acupuncture Inhibits Acetylcholine Synthesis and Restores Muscarinic Acetylcholine Receptor M2 Expression in the Lung When Treating Allergic Asthma Pubmed:29520557
International Journal of Molecular Sciences In Vitro Evaluation and Docking Studies of 5-oxo-5H-furo [3, 2-g] chromene-6-carbaldehyde Derivatives as Potential Anti-Alzheimer's Agents Pubmed: 31683761
BMC Anesthesiology Cerebrospinal fluid cholinergic biomarkers are associated with postoperative delirium in elderly patients undergoing Total hip/knee replacement: a … Pubmed: 32988385
PHYTOMEDICINE Thymoquinone administration ameliorates Alzheimer's disease-like phenotype by promoting cell survival in the hippocampus of amyloid beta1-42 infused rat … Pubmed: 32920292
PLoS One Neuroprotective effect of sodium alginate against chromium-induced brain damage in rats Pubmed:35421180
scientific reports Neuropsychopharmacological profiling of scoparone in mice Pubmed:35039558
SAUDI J BIOL SCI Aqueous Ajwa dates seeds extract improves memory impairment in type-2 diabetes mellitus rats by reducing blood glucose levels and enhancing brain … Pubmed:35531250
Saudi Pharmaceutical Journal Sukkari dates seed improves type-2 diabetes mellitus-induced memory impairment by reducing blood glucose levels and enhancing brain cholinergic … Pubmed:35812141
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