AffiASSAY® Lactate Colorimetric & Fluorometric Assay Kit
The AffiASSAY® Lactate Colorimetric & Fluorometric Assay Kit provides a reliable method for the quantification of lactate levels in various biological samples. Lactate is a key metabolite involved in cellular energy metabolism and is a critical marker in assessing lactate production and clearance in physiological and pathological conditions. This assay kit offers both colorimetric and fluorometric detection methods, providing researchers with flexibility and accuracy in measuring lactate concentrations.
Key Features:
- High Sensitivity: The assay kit can detect low levels of lactate, ensuring accurate quantification in biological samples with minimal interference.
- Flexible Detection: Offers both colorimetric and fluorometric detection methods, providing researchers with options based on their experimental needs and available instrumentation.
- Wide Application: Compatible with various sample types, including serum, plasma, tissue homogenates, and cell lysates, the AffiASSAY® Lactate Colorimetric & Fluorometric Assay Kit offers versatility in lactate-related research.
- Quick and Easy: With a streamlined protocol, you can obtain results rapidly, saving time and resources in the laboratory.
Assay Principle:
The assay kit measures lactate levels based on its enzymatic conversion to generate a product that reacts with a probe to produce a colorimetric or fluorometric signal. The intensity of the signal is directly proportional to the concentration of lactate in the sample, allowing for precise quantification.
Applications:
- Cellular Metabolism: Study lactate production and utilization in cellular energy metabolism and aerobic glycolysis.
- Exercise Physiology: Assess lactate kinetics and clearance during exercise and physical activity.
- Pathophysiology: Investigate the role of lactate in metabolic disorders, such as lactate acidosis, sepsis, and ischemia.
Kit Contents:
- Lactate Assay Buffer
- Lactate Enzyme Mix
- Lactate Probe
- Microplate (96 wells)
- 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® Lactate 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® Lactate Colorimetric & Fluorometric Assay Kit
- Biological samples containing lactate
- Microplate reader capable of measuring absorbance or fluorescence
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Standard Curve Preparation:
- Prepare a series of standard solutions containing known concentrations of lactate.
- Dilute the Lactate Assay Buffer and Lactate Enzyme Mix as instructed in the kit manual.
- Add the standard solutions to each well of the microplate to generate the standard curve.
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 lactate 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:
- Prepare the reaction mixture by combining the Lactate Assay Buffer and Lactate Enzyme Mix.
- Add the reaction mixture to each well of the microplate containing the samples and standards.
- Incubate the microplate at the appropriate temperature (as specified in the kit instructions) for a recommended time to allow the enzymatic conversion of lactate.
Colorimetric or Fluorometric Measurement:
- After the incubation period, measure the absorbance or fluorescence at the appropriate wavelength using a microplate reader.
Data Analysis:
- Calculate the lactate levels in your samples based on the absorbance or fluorescence readings and the standard curve generated from the standard solutions.
Results Interpretation:
- Interpret and analyze your results according to your research objectives.
- Observe any variations or trends in lactate 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® Lactate 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.