SKU: AFG-BV-0427

AffiASSAY® LDH Cytotoxicity Colorimetric Assay Kit

Vendor AffiGEN
Regular price 2.100,00 DKK
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AffiASSAY® LDH Cytotoxicity Colorimetric Assay Kit

The AffiASSAY® LDH Cytotoxicity Colorimetric Assay Kit provides a reliable method for the quantitative measurement of lactate dehydrogenase (LDH) release from damaged cells, allowing for the assessment of cytotoxicity in various biological samples. LDH is a cytoplasmic enzyme that is released into the extracellular space upon cell membrane damage, making it a useful marker for cytotoxicity evaluation. This assay kit utilizes a colorimetric detection method, enabling researchers to accurately assess cell viability and cytotoxic effects.

Key Features:

  • High Sensitivity: The assay kit can detect low levels of LDH release, ensuring accurate quantification of cytotoxicity in biological samples.
  • Wide Application: Compatible with various sample types, including cell culture supernatants, serum, plasma, and tissue homogenates, the AffiASSAY® LDH Cytotoxicity Colorimetric Assay Kit offers versatility in cytotoxicity assessment.
  • 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 release 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 of cytotoxicity.

Applications:

  • Cell Viability Assessment: Evaluate cell viability and cytotoxic effects of compounds, drugs, or treatments on various cell types.
  • Drug Screening: Assess the cytotoxicity of potential therapeutic agents or drug candidates in cell-based assays.
  • Environmental Toxicology: Investigate the cytotoxic effects of environmental pollutants or chemicals on cellular systems.

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 Cytotoxicity 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 Cytotoxicity Colorimetric Assay Kit
  • Cell culture supernatants or other biological samples
  • 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 release 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.
  • Assess the cytotoxic effects of compounds or treatments based on LDH release.

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 Cytotoxicity 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.