AffiASSAY® Aldehyde Dehydrogenase Activity Fluorometric Assay Kit
AffiASSAY® Aldehyde Dehydrogenase Activity Fluorometric Assay Kit is designed to provide precise and reliable measurements of aldehyde dehydrogenase (ALDH) activity in biological samples. This assay kit offers a fluorometric detection method, making it a valuable tool for researchers in the fields of biochemistry, cell biology, and molecular biology.
Key Features:
- High Sensitivity: Our assay kit is optimized to detect even low levels of ALDH activity, allowing for accurate quantification in a wide range of samples.
- Fluorometric Detection: Utilizing a highly sensitive fluorometric assay, our kit ensures robust and reproducible results, minimizing the margin for error.
- Wide Application: The AffiASSAY® Aldehyde Dehydrogenase Activity Fluorometric Assay Kit can be used with various sample types, including cell lysates, tissue homogenates, and purified enzyme preparations.
- Quick and Easy: With a simple and streamlined protocol, you can obtain results in just a few easy steps, saving you valuable time in the lab.
- Versatile: Whether you're studying cellular detoxification processes, investigating the effects of ALDH inhibitors, or screening potential therapeutic compounds, our kit provides the versatility you need.
Assay Principle:
This assay kit employs a fluorometric method based on the enzymatic conversion of aldehydes into their corresponding carboxylic acids by ALDH, coupled with the production of NADH. The increase in fluorescence intensity at specific wavelengths is directly proportional to the ALDH activity in your sample, allowing for precise quantification.
Applications:
- Detoxification Studies: Assess ALDH activity as a key enzyme involved in cellular detoxification processes, particularly in the metabolism of reactive aldehydes.
- Drug Screening: Screen potential ALDH inhibitors or activators for their pharmacological effects, particularly in the context of diseases associated with oxidative stress and aldehyde toxicity.
- Cellular Signaling: Investigate the role of ALDH in cellular signaling pathways and its implications in various physiological processes, including cell proliferation and apoptosis.
Kit Contents:
- Assay Buffer
- NAD+ (Nicotinamide Adenine Dinucleotide)
- Aldehyde Substrate
- ALDH Enzyme
- Positive Control
- 96-Well Assay Plate
- 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® Aldehyde Dehydrogenase Activity 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® Aldehyde Dehydrogenase Activity Fluorometric Assay Kit
- Biological samples containing ALDH
- 96-well microplate
- Microplate reader capable of measuring fluorescence intensity at specific wavelengths
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Sample Preparation:
- Prepare your biological samples containing ALDH. These samples can be cell lysates, tissue homogenates, or purified enzyme preparations.
- Ensure that your samples are stored and handled according to your specific experimental requirements.
Assay Plate Setup:
- Label the microplate wells according to your experimental design, including wells for your samples, standards, and controls.
- If using controls, prepare a positive control sample with known ALDH activity.
Assay Reaction:
- Prepare the assay reaction mix by combining the Assay Buffer, NAD+, Aldehyde Substrate, and ALDH Enzyme from the kit as instructed.
- Add 50-100 µL of the assay reaction mix to each well of the microplate.
Sample and Control Addition:
- Add 50-100 µL of your prepared samples and positive control to the appropriate wells on the microplate.
- Ensure that the final reaction volume in each well is consistent.
Incubation:
- Incubate the microplate at the appropriate temperature (as specified in the kit instructions) for a recommended time to allow the enzymatic reaction to occur.
Fluorescence Measurement:
- Using a microplate reader capable of measuring fluorescence intensity at specific wavelengths, measure the fluorescence intensity of each well.
- Record the fluorescence emission at the appropriate excitation and emission wavelengths.
Data Analysis:
- Calculate the ALDH activity in your samples based on their respective fluorescence intensity readings.
- Compare the activity levels to your positive control and perform any necessary normalization.
Results Interpretation:
- Interpret and analyze your results according to your research objectives.
- Note any variations or trends in ALDH 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® Aldehyde Dehydrogenase Activity 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.