SKU: AFG-BV-0697

AffiASSAY® LDH Activity Colorimetric Assay Kit

Vendor AffiGEN
Regular price £303.00 GBP
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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.