AffiASSAY® Glutathione Peroxidase Activity Colorimetric Assay Kit
The AffiASSAY® Glutathione Peroxidase Activity Colorimetric Assay Kit offers a reliable method for the quantification of glutathione peroxidase (GPx) activity in biological samples. This assay kit provides a colorimetric detection method, enabling researchers to study antioxidant enzyme activity, oxidative stress, and redox signaling with precision and convenience.
Key Features:
- High Sensitivity: The assay kit can detect low levels of GPx activity, ensuring accurate quantification in biological samples.
- Wide Application: The AffiASSAY® Glutathione Peroxidase Activity Colorimetric Assay Kit is compatible with various sample types, including cell lysates, tissue homogenates, and plasma.
- 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 GPx activity, reducing experimental costs without compromising data quality.
Assay Principle:
The assay kit measures GPx activity based on the reduction of hydroperoxide by GPx, coupled with the oxidation of glutathione (GSH) to oxidized glutathione (GSSG) by GPx. The decrease in absorbance at 340 nm is directly proportional to GPx activity in the sample.
Applications:
- Antioxidant Research: Investigate the role of GPx in protecting cells from oxidative damage and maintaining cellular redox homeostasis.
- Pathophysiology Studies: Study the involvement of GPx activity in various diseases, including cancer, neurodegenerative disorders, and cardiovascular diseases.
- Drug Discovery: Screen for compounds that modulate GPx activity as potential therapeutics for oxidative stress-related diseases.
Kit Contents:
- GPx Assay Buffer
- GPx Substrate
- GPx Enzyme
- Glutathione Reductase (GR)
- NADPH
- Microplate (96 wells)
- GPx Standard
- 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® Glutathione Peroxidase Activity Colorimetric 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® Glutathione Peroxidase Activity Colorimetric Assay Kit
- Biological samples containing GPx
- Microplate reader capable of measuring absorbance at 340 nm
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Standard Curve Preparation:
- Prepare a series of standard solutions containing known concentrations of the GPx standard provided in the kit.
- Dilute the GPx Substrate and GPx Enzyme 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 GPx 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 GPx Assay Buffer, GPx Substrate, GPx Enzyme, Glutathione Reductase (GR), and NADPH.
- 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 Measurement:
- After the incubation period, measure the absorbance at 340 nm using a microplate reader.
Data Analysis:
- Calculate the GPx activity in your samples based on the absorbance readings and 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 GPx activity 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® Glutathione Peroxidase Activity Colorimetric 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.