AffiASSAY® Choline/Acetylcholine Colorimetric & Fluorometric Assay Kit
AffiASSAY® Choline/Acetylcholine Colorimetric & Fluorometric Assay Kit is designed to provide precise and reliable measurements of choline and acetylcholine levels in biological samples. This assay kit offers both colorimetric and fluorometric detection methods, making it a versatile tool for researchers in neuroscience, pharmacology, and neurology.
Key Features:
- High Sensitivity: Our assay kit is optimized to detect choline and acetylcholine levels with high sensitivity, allowing for accurate quantification even in low concentration samples.
- Colorimetric & Fluorometric Detection: With the flexibility to choose between colorimetric or fluorometric detection methods, our kit provides options for different experimental setups and preferences.
- Wide Application: The AffiASSAY® Choline/Acetylcholine Assay Kit can be used with various sample types, including brain tissue, serum, plasma, and cell lysates.
- 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 cholinergic neurotransmission, investigating the effects of drugs on acetylcholine levels, or exploring choline metabolism, our kit provides the versatility you need.
Assay Principle:
This assay kit measures choline and acetylcholine levels based on the enzymatic reactions involving choline and acetylcholine. Choline is oxidized to betaine, releasing hydrogen peroxide in the process, which reacts with a colorimetric or fluorometric probe to generate a signal proportional to choline concentration. Acetylcholine is hydrolyzed to choline and acetate by acetylcholinesterase, and the released choline is then detected using the same enzymatic reaction as for choline measurement.
Applications:
- Neuroscience: Study cholinergic neurotransmission and its role in cognitive function, learning, and memory.
- Pharmacology: Assess the effects of drugs targeting cholinergic receptors or acetylcholinesterase inhibitors.
- Neurology: Investigate choline and acetylcholine levels as potential biomarkers for neurological disorders such as Alzheimer's disease.
Kit Contents:
- Choline/Acetylcholine Assay Buffer
- Choline/Acetylcholine Enzyme Mix
- Choline Probe
- Acetylcholine Probe
- 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® Choline/Acetylcholine 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® Choline/Acetylcholine Colorimetric & Fluorometric Assay Kit
- Biological samples containing choline and/or acetylcholine
- 96-well microplate
- Microplate reader capable of measuring absorbance or fluorescence intensity
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Standard Curve Preparation:
- Prepare a series of standard solutions containing known concentrations of choline and/or acetylcholine.
- Dilute the Choline/Acetylcholine Probe provided in the kit with the Assay Buffer to prepare the working solution.
- Add the working solution to each standard solution to initiate the reaction.
Assay Plate Setup:
- Label the microplate wells according to your experimental design, including wells for your samples, standards, and controls.
- If using controls, prepare control samples without choline or acetylcholine to serve as background controls.
Sample and Standard Addition:
- Add 50-100 µL of your prepared samples and standard solutions to the appropriate wells on the microplate.
- Ensure that the final reaction volume in each well is consistent.
Assay Reaction:
- Add 50 µL of Choline/Acetylcholine Enzyme Mix to each well to initiate the reaction.
- Incubate the microplate at the appropriate temperature (as specified in the kit instructions) for a recommended time to allow the enzymatic reactions to occur.
Signal Detection:
- For colorimetric detection: Measure the absorbance at the appropriate wavelength using a microplate reader.
- For fluorometric detection: Measure the fluorescence intensity using a microplate reader with excitation and emission wavelengths suitable for the probes provided in the kit.
Data Analysis:
- Generate a standard curve using the absorbance or fluorescence values obtained from the standard solutions.
- Calculate the choline and/or acetylcholine concentrations in your samples based on their respective absorbance or fluorescence readings and the standard curve.
Results Interpretation:
- Interpret and analyze your results according to your research objectives.
- Note any variations or trends in choline and/or acetylcholine 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® Choline/Acetylcholine 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.