SKU: AFG-BV-0041

AffiASSAY® Alanine Aminotransferase (ALT or SGPT) Activity Colorimetric & Fluorometric Assay Kit

Vendor AffiGEN
Regular price $380.00 USD
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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.