ELISA Kit for Acetylcholinesterase (ACHE)

ARAChE; N-AChE; YT

Specificity

This assay has high sensitivity and excellent specificity for detection of Acetylcholinesterase (ACHE).
No significant cross-reactivity or interference between Acetylcholinesterase (ACHE) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 102
EDTA plasma(n=5) 78-97 85
heparin plasma(n=5) 93-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Acetylcholinesterase (ACHE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Acetylcholinesterase (ACHE) 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) 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) 82-92% 78-94% 79-96% 78-96%
EDTA plasma(n=5) 86-93% 97-105% 95-102% 96-104%
heparin plasma(n=5) 95-103% 81-96% 78-103% 89-99%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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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