AffiASSAY® LDH Activity Colorimetric Assay Kit
The AffiASSAY® LDH Activity Colorimetric Assay Kit offers a reliable method for the quantitative measurement of lactate dehydrogenase (LDH) activity in various biological samples. LDH is a key enzyme involved in cellular metabolism, catalyzing the interconversion of lactate and pyruvate. This assay kit utilizes a colorimetric detection method, enabling researchers to accurately assess LDH activity with ease.
Key Features:
- High Sensitivity: The assay kit can detect low levels of LDH activity, ensuring accurate quantification in biological samples.
- Wide Application: Compatible with various sample types, including serum, plasma, tissue homogenates, and cell lysates, the AffiASSAY® LDH Activity Colorimetric Assay Kit offers versatility in LDH-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 LDH activity based on its enzymatic conversion of lactate to pyruvate, coupled with the reduction of a colorimetric probe. The intensity of the colorimetric signal is directly proportional to the LDH activity in the sample, allowing for precise quantification.
Applications:
- Cellular Metabolism: Study LDH activity in cellular energy metabolism and glycolysis.
- Tissue Damage Assessment: Assess LDH release as a marker of cellular damage or necrosis in various pathophysiological conditions.
- Drug Toxicity Screening: Evaluate the cytotoxic effects of drugs or chemicals on cells based on LDH release.
Kit Contents:
- LDH Assay Buffer
- LDH Substrate Mix
- LDH Enzyme Mix
- LDH Positive Control
- 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® LDH Activity Colorimetric 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® LDH Activity Colorimetric Assay Kit
- Biological samples containing LDH
- Microplate reader capable of measuring absorbance
- Pipettes and tips
- Microcentrifuge tubes
Protocol:
Standard Curve Preparation:
- Prepare a series of standard solutions containing known concentrations of LDH positive control.
- Dilute the LDH Assay Buffer, LDH Substrate Mix, and LDH 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 LDH 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 LDH Assay Buffer, LDH Substrate Mix, and LDH 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 Measurement:
- After the incubation period, measure the absorbance at the appropriate wavelength using a microplate reader.
Data Analysis:
- Calculate the LDH activity in your samples based on the absorbance 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 LDH 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® LDH Activity Colorimetric 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.