AffiASSAY® Alanine Aminotransferase (ALT or SGPT) Activity Colorimetric & Fluorometric Assay Kit
AffiASSAY® Alanine Aminotransferase (ALT or SGPT) Activity Colorimetric & Fluorometric Assay Kit is designed to provide precise and reliable measurements of ALT activity in biological samples. This assay kit offers both colorimetric and fluorometric detection methods, making it a versatile 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 ALT activity, allowing for accurate quantification in a wide range of samples.
- Colorimetric and Fluorometric Detection: With both colorimetric and fluorometric detection options, our kit ensures flexibility and convenience, catering to different experimental needs.
- Wide Application: The AffiASSAY® Alanine Aminotransferase (ALT or SGPT) Activity 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 liver function, investigating disease mechanisms, or screening potential therapeutics, our kit provides the versatility you need.
Assay Principle:
This assay kit employs a colorimetric or fluorometric method based on the enzymatic conversion of alanine and α-ketoglutarate into pyruvate and glutamate by ALT. The generated product can be detected either through colorimetric changes or fluorescence emission, allowing for precise quantification of ALT activity.
Applications:
- Liver Function Studies: Assess ALT activity as a biomarker of liver health and function.
- Disease Mechanism Investigations: Investigate the role of ALT in various diseases, including liver diseases and metabolic disorders.
- Drug Screening: Screen potential therapeutics for their impact on ALT activity, aiding in drug development.
- Cellular Metabolism: Study the involvement of ALT in cellular metabolism and amino acid metabolism.
Kit Contents:
- Assay Buffer
- ALT Enzyme
- Developer Solution
- Substrate Mix
- 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® Alanine Aminotransferase (ALT or SGPT) Activity 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® Alanine Aminotransferase (ALT or SGPT) Activity Colorimetric & Fluorometric Assay Kit
- Biological samples containing ALT
- 96-well microplate
- Microplate reader capable of measuring absorbance or fluorescence
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Sample Preparation:
- Prepare your biological samples containing ALT. 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 ALT activity.
Assay Reaction:
- Prepare the assay reaction mix by combining the Assay Buffer, ALT Enzyme, and Substrate Mix 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.
Measurement:
- Using a microplate reader capable of measuring absorbance or fluorescence, measure the absorbance or fluorescence intensity of each well.
- For colorimetric detection, measure the absorbance at the appropriate wavelength.
- For fluorometric detection, measure the fluorescence emission at the appropriate excitation and emission wavelengths.
Data Analysis:
- Calculate the ALT activity in your samples based on their respective absorbance or fluorescence 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 ALT 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® Alanine Aminotransferase (ALT or SGPT) Activity 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.