AffiASSAY® Total Polyamine Fluorometric Assay Kit
Unlock the mysteries of polyamines with the AffiASSAY® Total Polyamine Fluorometric Assay Kit. This innovative kit provides precise and reliable measurements of total polyamine levels across a variety of biological samples, empowering researchers in biochemistry, cell biology, and molecular biology to make groundbreaking discoveries.
Key Features:
- Unmatched Sensitivity: Our cutting-edge assay kit is fine-tuned to detect even the faintest levels of polyamines, ensuring accurate quantification in diverse samples.
- Fluorometric Brilliance: Harnessing a highly sensitive fluorometric assay, our kit guarantees robust and reproducible results, reducing the margin for error.
- Broad Applicability: The AffiASSAY® Total Polyamine Fluorometric Assay Kit is versatile enough to handle a wide range of sample types, from cells and tissues to purified enzyme preparations.
- Efficiency at Its Best: With a streamlined protocol, you can achieve results in just a few straightforward steps, saving precious time in the lab.
- Research Versatility: Whether delving into cellular metabolism, assessing drug impacts, or probing enzymatic reactions, our kit provides the flexibility and precision you need.
Assay Principle:
Our assay kit leverages a fluorometric method based on the enzymatic conversion of polyamines into a fluorescent product. The increase in fluorescence intensity is directly proportional to the polyamine concentration in your sample, enabling precise quantification.
Applications:
- Metabolic Research: Investigate the role of polyamines in cellular growth, differentiation, and energy production.
- Drug Development: Evaluate the effects of pharmaceutical compounds on polyamine levels, facilitating drug discovery and development.
- Enzyme Kinetics: Explore the kinetics of polyamine-utilizing enzymes with precision and ease.
- Cellular Signaling: Delve into the intricate connections between polyamines and cellular signaling pathways.
Kit Contents:
- Assay Buffer
- Polyamine Standard
- Enzyme Mix
- Developer Solution
- 96-Well Assay Plate
- Comprehensive Protocol
Why Choose AffiASSAY®?
At AffiASSAY®, we are dedicated to excellence and scientific rigor. Our commitment to providing innovative tools and precise results makes us the trusted choice of researchers worldwide. Experience the AffiASSAY® difference and accelerate your research with confidence.
AffiASSAY® Total Polyamine 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® Total Polyamine Fluorometric Assay Kit
- Biological samples containing polyamines
- 96-well microplate
- Fluorescence microplate reader capable of measuring at the appropriate excitation/emission wavelengths
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Sample Preparation:
- Prepare your biological samples containing polyamines. 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 Polyamine Standard provided in the kit according to the kit's instructions.
- Create a series of standard dilutions using the reconstituted Polyamine 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 polyamines 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 polyamine 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 Polyamine 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 polyamines to a fluorescent product.
Fluorescence Measurement:
- Using a fluorescence microplate reader, measure the fluorescence intensity of each well. The specific excitation/emission wavelengths will depend on the fluorometric detection method used.
Data Analysis:
- Generate a standard curve using the fluorescence values obtained from the Polyamine Standards.
- Calculate the polyamine 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 polyamine 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® Total Polyamine 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.