AffiASSAY® Acetyl-CoA Fluorometric Assay Kit

AffiASSAY® Acetyl-CoA Fluorometric Assay Kit

AffiASSAY® Acetyl-CoA Fluorometric Assay Kit is designed to provide precise and reliable measurements of Acetyl-CoA levels in a variety of biological samples, this assay kit is a powerful 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 Acetyl-CoA, 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® Acetyl-CoA Fluorometric Assay Kit can be used with a broad range of sample types, including cells, tissues, 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 metabolism, investigating the effects of drug compounds, or exploring the intricacies of enzymatic reactions, our kit provides the versatility you need.

Assay Principle:

This assay kit employs a fluorometric method based on the enzymatic conversion of Acetyl-CoA into a fluorescent product. The increase in fluorescence intensity is directly proportional to the Acetyl-CoA concentration in your sample, allowing for precise quantification.

Applications:

• Metabolic Research: Study the role of Acetyl-CoA in cellular energy production and lipid metabolism.
• Drug Development: Assess the impact of pharmaceutical compounds on Acetyl-CoA levels, aiding in drug discovery.
• Enzyme Kinetics: Investigate the kinetics of Acetyl-CoA-utilizing enzymes with accuracy and ease.
• Cellular Signaling: Explore the connections between Acetyl-CoA and cellular signaling pathways.

Kit Contents:

• Assay Buffer
• Acetyl-CoA Standard
• Enzyme Mix
• Developer Solution
• 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® Acetyl-CoA 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® Acetyl-CoA Fluorometric Assay Kit
• Biological samples containing Acetyl-CoA
• 96-well microplate
• Microplate reader capable of measuring fluorescence (excitation: ~535-575 nm, emission: ~585-625 nm)
• Pipettes and tips
• Microcentrifuge tubes

Protocol:

Sample Preparation:

• Prepare your biological samples containing Acetyl-CoA. 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.

Standard Curve Preparation:

• Reconstitute the Acetyl-CoA Standard provided in the kit according to the kit's instructions.
• Create a series of standard dilutions using the reconstituted Acetyl-CoA standard to generate a standard curve. Typical concentrations may range from 0 to 100 µM.

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 control sample without Acetyl-CoA to serve as a background control.

Sample Dilution:

• Dilute your biological samples as needed to ensure that they fall within the linear range of the standard curve. The specific dilution factor will depend on the expected Acetyl-CoA concentration in your samples.

Assay Reaction:

• Prepare the assay reaction mix by combining the Assay Buffer and Enzyme Mix from the kit as instructed.
• Add 50-100 µL of the assay reaction mix to each well of the microplate.

Sample and Standard Addition:

• Add 50-100 µL of your diluted samples and Acetyl-CoA standards 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 conversion of Acetyl-CoA to a fluorescent product.

Fluorescence Measurement:

• Using a microplate reader with appropriate fluorescence settings, measure the fluorescence intensity of each well. Excitation should be around 535-575 nm, and emission around 585-625 nm.

Data Analysis:

• Generate a standard curve using the fluorescence values obtained from the Acetyl-CoA standards.
• Calculate the Acetyl-CoA concentrations in your samples based on their respective fluorescence readings and the standard curve.

Results Interpretation:

• Interpret and analyze your results according to your research objectives.
• Note any variations or trends in Acetyl-CoA levels 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® Acetyl-CoA 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.