ELISA Kit for Beta-Site APP Cleaving Enzyme 1 (bACE1)

ASP2; Beta Secretase; Memapsin 2; Aspartyl protease 2; Beta-site amyloid precursor protein cleaving enzyme 1; Membrane-associated aspartic protease 2

Specificity

This assay has high sensitivity and excellent specificity for detection of Beta-Site APP Cleaving Enzyme 1 (bACE1).
No significant cross-reactivity or interference between Beta-Site APP Cleaving Enzyme 1 (bACE1) and analogues was observed.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Beta-Site APP Cleaving Enzyme 1 (bACE1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Beta-Site APP Cleaving Enzyme 1 (bACE1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

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
Food & Function Comparative study of the effects of phosphatidylcholine rich in DHA and EPA on Alzheimer’s disease and the possible mechanisms in CHO-APP/PS1 cells and SAMP8 mice pubmed:29292421
Journal of Functional Foods EPA enriched ethanolamine plasmalogens significantly improve cognition of Alzheimer's disease mouse model by suppressing β-amyloid generation 10.1016/j.jff.2017.12.016
Journal of Nutritional Biochemistry Mechanisms of DHA-enriched phospholipids in improving cognitive deficits in aged SAMP8 mice with high-fat diet Pubmed:29986309
Neurotoxicology Involvement of hippocampal agmatine in β1-42 amyloid induced memory impairment, neuroinflammation and BDNF signaling disruption in mice Pubmed: 32522471
Food Chem Toxicol Effects of Alcohol Intake on Cognitive Function and ¦Â-amyloid Protein in APP/PS1 Transgenic Mice 33737111
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