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General Adenosine Triphosphate (ATP) ELISA Kit

SKU: orb781738
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Description

General Adenosine Triphosphate (ATP) ELISA Kit

Research Area

Pharmacology & Drug Discovery

Images & Validation

Tested ApplicationsELISA
Application Notes
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with ATP. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to ATP. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of ATP in the samples is then determined by comparing the OD of the samples to the standard curve.

Key Properties

ReactivityAll
Sample Typesserum, plasma, cell lysates, cell culture supernates and other biological fluids
Assay TypeCompetitive
Assay Time2h
Range15.63-1000 ng/mL
Sensitivity5.18 ng/mL
Concentration1000 ng/mL

Procedure & Performance

Assay Principle
This kit is based on a competitive ELISA detection method. The microtiter plate is pre-coated with target antigen. During the assay, the target antigen in the sample or standard competes with the immobilized target for binding sites on the biotinylated detection antibody specific to the target antigen. After incubation, excess conjugate and unbound sample or standard are removed by washing. HRP–streptavidin (SABC) is then added to each well and incubated, followed by the addition of TMB substrate solution. The enzymatic reaction is terminated by adding an acidic stop solution, and the color change is measured at 450 nm using a microplate reader. The concentration of the target antigen in the samples is determined by comparison with the standard curve and is inversely proportional to the OD450 value.
Kit Components
1. ELISA Microplate
2. Standards
3. Detection Antibody
4. HRP-Streptavidin Conjugate
5. TMB Substrate
6. Dilution buffers
7. Stop Solution
8. Wash Buffer
9. Plate Sealers
10. Manual
Reagent Preparation
1. Wash Buffer: Prepare the 1X Wash Buffer using distilled water according to the manual.
2. Standard: Perform gradient dilution according to the instructions in the manual.
3. Other Concentrated Reagents: Dilute the concentrated reagents using the Dilution Buffers provided in the kit to 1 X working solutions as instructed in the manual. Always use a clean pipette tip for each different solution.
Assay Procedure
This procedure is for reference only.

1. After the kit equilibrates to room temperature, add standards or samples to each well and incubate.
2. Add biotinylated antibody working solution to each well and incubate.
3. Discard liquid, add wash buffer to each well, wash the plate three times, and blot dry on clean absorbent paper.
4. Add streptavidin-HRP working solution to each well and incubate._x000b_5. Discard liquid, add wash buffer to each well, wash the plate five times, and blot dry on clean absorbent paper.
6. Add TMB substrate solution to each well and incubate in the dark.
7. Add stop solution to each well, mix thoroughly, and immediately read OD at 450 nm.
Materials Required
1. Microplate readers
2. Centrifuge
3. Incubator
4. Automated plate washer
5. Single-channel or multi-channel high-precision pipettes
6. Disposable pipette tips
7. Sterile tubes
8. Eppendorf tubes
9. Absorbent paper
10. Loading slots
Precision
Intra-assay Precision (Precision within an assay): CV% < 8%
Intra-assay precision was evaluated by testing multiple replicates of samples within the same plate.

Inter-assay Precision (precision between assays): CV% < 10%
Inter-assay precision was evaluated by testing samples across different plates.
Calculation of Results
1. Average the duplicate readings for each Standard, Control, and Sample, and subtract the mean optical density of the zero Standard.
2. Construct a standard curve by plotting the target concentration on the y-axis against absorbance on the x-axis and draw a curve through the data points.
3. Determine the sample concentration by substituting the OD450 value into the standard curve. For diluted samples, multiply the calculated value by the corresponding dilution factor.
Curve Fitting Softwares
1. Curve Expert
2. Thermo SkanIt RE
3. SciDAVis
4. LabPlot
5. ……

Storage & Handling

StorageRefer to the Storage Guidelines in the Manual
Expiration DatePlease enquire.
DisclaimerFor research use only

Alternative Names

Adenosine-5'-Triphosphate ELISA Kit
Quality Guarantee

Quality Guarantee

Explore bioreagents carefree to elevate your research. All our products are rigorously tested for performance. If a product does not perform as described on its datasheet, our scientific support team will provide expert troubleshooting, a prompt replacement, or a refund. For full details, please see our Terms & Conditions and Buying Guide. Contact us at support@biorbyt.com.

General Adenosine Triphosphate (ATP) ELISA Kit

Standard curve for orb781738 ELISA kit.

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Datasheet
Product Information
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Manual
Product Manual
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Protocol Information

ELISA
Enzyme-linked Immunosorbent Assay (EIA)
View Protocol

General Adenosine Triphosphate (ATP) ELISA Kit (orb781738)

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Applications: ELISA|Reactivity: Mouse

I recently used Biorbyt’s Universal ATP ELISA Kit in our laboratory and was very satisfied with the overall experience. I highly recommend it to researchers who need quantitative ATP detection. Advantages: Simple and convenient operation: The kit uses a competitive inhibition enzyme immunoassay method with a clear and straightforward workflow. The assay can be completed in approximately 2 hours, making it suitable for routine experiments. The pre-coated ATP plate eliminates many preparation steps. Suitable sensitivity and detection range: The detection range is 15.63–1000 ng/mL, with a sensitivity of approximately 5.18 ng/mL. It effectively covers the ATP concentration requirements of most biological samples, including serum, plasma, cell lysates, and cell culture supernatants. Good reproducibility: The standard curve showed good linearity (the attached figure demonstrates a smooth standard curve), and the CV values between replicate wells were low, indicating reliable and consistent results. Broad applicability: As a universal kit suitable for samples from multiple species, it is highly practical for research areas such as pharmacology, drug discovery, and cellular metabolism. Good cost-effectiveness: Compared with similar products, Biorbyt’s kit is competitively priced and is available in both 48T and 96T formats, making it suitable for experiments of different scales. User experience: In experiments related to cellular energy metabolism, this kit showed stable performance, low background, clear color development, and consistent absorbance readings at 450 nm. The instruction manual was detailed, and technical support was responsive (although I did not encounter any issues during this experiment). Our laboratory is planning to continue purchasing this product. If you are conducting ATP quantification studies, this kit is a reliable choice.

- Li ZhangHefei University of Technology / Pharmacology
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Applications: Bioassay, ELISA|Reactivity: Mouse

I recently used Biorbyt’s Universal ATP ELISA Kit in our laboratory and was very satisfied with the overall performance. I highly recommend it to researchers who need quantitative ATP detection. Advantages: Simple and convenient operation: The kit uses a competitive inhibition enzyme immunoassay method with a clear and straightforward workflow. The assay can be completed in approximately 2 hours, making it suitable for routine experiments. The ATP-coated plate eliminates the need for extensive preparation steps. Suitable sensitivity and detection range: The detection range is 15.63–1000 ng/mL, with a sensitivity of approximately 5.18 ng/mL, covering the ATP concentration requirements of most biological samples, including serum, plasma, cell lysates, and cell culture supernatants. Good reproducibility: The standard curve showed good linearity (the attached figure demonstrates a smooth standard curve), and the CV values between replicate wells were low, indicating reliable results. Broad applicability: As a universal kit suitable for samples from multiple species, it is highly useful for research areas such as pharmacology, drug discovery, and cellular metabolism. Good cost performance: Compared with similar products, Biorbyt’s kit is competitively priced and is available in both 48T and 96T formats, making it suitable for experiments of different scales. User experience: In cellular energy metabolism studies, this kit showed stable performance, low background, clear color development, and consistent absorbance readings at 450 nm. The instruction manual was detailed, and technical support was responsive (although I did not encounter any issues during this experiment). Our laboratory is planning to continue purchasing this product. For researchers conducting ATP quantification studies, this kit is a reliable choice.

- Harry FangHefei University of technology / Pharmacology

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48 T
$ 350.00
96 T
$ 490.00
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