AffiASSAY® Alpha Galactosidase (α-Gal) Activity Fluorometric Assay Kit
AffiASSAY® Alpha Galactosidase (α-Gal) Activity Fluorometric Assay Kit is designed to provide precise and reliable measurements of alpha galactosidase 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 α-Gal 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® Alpha Galactosidase 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 lysosomal storage disorders, investigating carbohydrate metabolism, 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 hydrolysis of a synthetic fluorogenic substrate by α-Gal, leading to the release of a fluorescent product. The increase in fluorescence intensity at specific wavelengths is directly proportional to the α-Gal activity in your sample, allowing for precise quantification.
Applications:
- Lysosomal Storage Disorders: Measure α-Gal activity as a diagnostic marker for lysosomal storage disorders, such as Fabry disease.
- Carbohydrate Metabolism: Study the role of α-Gal in carbohydrate metabolism and glycosphingolipid degradation pathways.
- Drug Screening: Screen potential α-Gal inhibitors or activators for their pharmacological effects, particularly in the context of lysosomal storage disorders.
Kit Contents:
- Assay Buffer
- Fluorogenic Substrate
- α-Gal Enzyme
- Positive Control
- Fluorescence Enhancer
- 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® Alpha Galactosidase (α-Gal) 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® Alpha Galactosidase (α-Gal) Activity Fluorometric Assay Kit
- Biological samples containing α-Gal
- 96-well microplate
- Fluorescence microplate reader capable of measuring fluorescence intensity at specific wavelengths
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Sample Preparation:
- Prepare your biological samples containing α-Gal. 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 α-Gal activity.
Assay Reaction:
- Prepare the assay reaction mix by combining the Assay Buffer, Fluorogenic Substrate, α-Gal Enzyme, and Fluorescence Enhancer 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 fluorescence microplate reader capable of measuring fluorescence intensity at specific wavelengths (excitation and emission wavelengths provided by the kit), measure the fluorescence intensity of each well.
Data Analysis:
- Calculate the α-Gal 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 α-Gal 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® Alpha Galactosidase (α-Gal) 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.
