AffiASSAY® Hydrogen Peroxide Colorimetric & Fluorometric Assay Kit
The AffiASSAY® Hydrogen Peroxide Colorimetric & Fluorometric Assay Kit offers a reliable method for the quantification of hydrogen peroxide levels in various biological samples. This versatile assay kit provides both colorimetric and fluorometric detection methods, allowing researchers to study oxidative stress, enzymatic reactions, and cellular signaling pathways with precision and flexibility.
Key Features:
- Dual Detection Methods: The kit offers both colorimetric and fluorometric assays, providing flexibility in detecting hydrogen peroxide levels based on experimental requirements.
- High Sensitivity: The assay kit can detect low levels of hydrogen peroxide, ensuring accurate quantification in biological samples.
- Wide Application: Compatible with various sample types, including cell lysates, tissue homogenates, and biological fluids, the AffiASSAY® Hydrogen Peroxide Colorimetric & Fluorometric Assay Kit offers versatility in oxidative stress studies.
- Quick and Easy: With a streamlined protocol, you can obtain results rapidly, saving time and resources in the laboratory.
- Cost-Effective: Our kit provides an affordable solution for measuring hydrogen peroxide levels, reducing experimental costs without compromising data quality.
Assay Principle:
The assay kit measures hydrogen peroxide levels based on the enzymatic reaction involving horseradish peroxidase (HRP), which oxidizes the substrate to produce a colored or fluorescent product. The intensity of the color or fluorescence is directly proportional to the concentration of hydrogen peroxide in the sample.
Applications:
- Oxidative Stress Research: Investigate the role of hydrogen peroxide in oxidative stress-related diseases and cellular damage.
- Enzymatic Reaction Studies: Assess the activity of enzymes involved in hydrogen peroxide metabolism and signaling pathways.
- Cell Signaling Pathways: Study the involvement of hydrogen peroxide in cellular signaling events and redox regulation.
Kit Contents:
- Hydrogen Peroxide Assay Buffer
- Hydrogen Peroxide Substrate
- Hydrogen Peroxide Enzyme Mix
- Microplate (96 wells)
- Detailed Protocol
Why Choose AffiASSAY®?
Our commitment to excellence and scientific rigor makes AffiASSAY® the trusted choice of researchers worldwide. We strive to provide you with innovative tools that deliver precise results and accelerate your research efforts.
AffiASSAY® Hydrogen Peroxide Colorimetric & Fluorometric Assay Kit Protocol:
[Please note that this protocol is a general guideline, and you should always refer to the specific instructions provided in the kit manual for accurate and detailed information.]
Materials Needed:
- AffiASSAY® Hydrogen Peroxide Colorimetric & Fluorometric Assay Kit
- Biological samples containing hydrogen peroxide
- Microplate reader capable of measuring absorbance or fluorescence
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Standard Curve Preparation:
- Prepare a series of standard solutions containing known concentrations of hydrogen peroxide.
- Dilute the Hydrogen Peroxide Assay Buffer, Hydrogen Peroxide Substrate, and Hydrogen Peroxide Enzyme Mix as instructed in the kit manual.
- Add the standard solutions to each well of the microplate to generate the standard curve.
Assay Plate Setup:
- Label the microplate wells according to your experimental design, including wells for your samples, standards, and controls.
- If using controls, prepare control samples without hydrogen peroxide to serve as background controls.
Sample and Standard Addition:
- Add 50-100 µL of your prepared samples and standard solutions to the appropriate wells on the microplate.
- Ensure that the final reaction volume in each well is consistent.
Assay Reaction:
- Prepare the reaction mixture by combining the Hydrogen Peroxide Assay Buffer, Hydrogen Peroxide Substrate, and Hydrogen Peroxide Enzyme Mix.
- Add the reaction mixture to each well of the microplate containing the samples and standards.
- Incubate the microplate at the appropriate temperature (as specified in the kit instructions) for a recommended time to allow the enzymatic reaction to proceed.
Colorimetric or Fluorometric Measurement:
- After the incubation period, measure the absorbance or fluorescence at the appropriate wavelength using a microplate reader.
Data Analysis:
- Calculate the hydrogen peroxide levels in your samples based on the absorbance or fluorescence readings and the standard curve generated from the standard solutions.
Results Interpretation:
- Interpret and analyze your results according to your research objectives.
- Observe any variations or trends in hydrogen peroxide levels among your samples.
Data Presentation:
- Prepare your data for presentation or publication, and include the appropriate statistical analysis if required.
Cleanup:
- Dispose of all waste materials, including used assay components, according to your laboratory's safety protocols.
- Always refer to the specific instructions provided with the AffiASSAY® Hydrogen Peroxide Colorimetric & Fluorometric Assay Kit for any kit-specific details or variations in the protocol.
- Additionally, ensure that you adhere to the best practices and safety guidelines in your laboratory while conducting the assay.