ELISA Kit for Adenosine Triphosphate (ATP)

Adenosine-5'-Triphosphate

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 87-95 91
EDTA plasma(n=5) 89-101 96
heparin plasma(n=5) 97-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Adenosine Triphosphate (ATP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Adenosine Triphosphate (ATP) 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 Adenosine Triphosphate (ATP) 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) 84-101% 87-102% 87-95% 88-102%
EDTA plasma(n=5) 85-103% 80-90% 96-104% 78-101%
heparin plasma(n=5) 98-105% 90-103% 80-94% 78-94%

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 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Iranian Journal of Health and Physical activity Effects of Aerobic Training, with or without Zizyphus Jujuba Water Extraction, on Fundus Nesfatin-1, ATP, HDL-C, and LDL-C Concentrations in Female Rats Ijvst: Source
Journal of Neurochemistry Neuroprotective effects of vildagliptin in rat rotenone Parkinson&#039;s disease model: role of RAGE‐NFκB and Nrf2‐antioxidant signaling pathways PubMed: 25752913
American Journal of Animal and Veterinary Sciences Metabolic Features of Heart Failure with Different Etiology ofsp11075
toxicology in vitro Rifampicin-induced injury in L02 cells is alleviated by 4-PBA via inhibition of the PERK-ATF4-CHOP pathway. pubmed:27470132
Free radical biology and medicine Rifampicin-induced injury in HepG2 cells is alleviated by TUDCA via increasing bile acid transporters expression and enhancing the Nrf2-mediated adaptive response. pubmed:28688954
Dipòsit Digital de la Universitat de Barcelona Chemiluminescent bioanalytical assays for clinical biomarkers 2445/116053
Evidence-Based Complementary and Alternative Medicine The Antioxidative Action of ZTP by Increasing Nrf2/ARE Signal Pathway
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