AffiASSAY® Cell Counting Kit-8
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Size: 3000 Assays

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AffiASSAY® Cell Counting Kit-8 (WST-8 / CCK-8)

Available Sizes: 1 ml, 2 x 1 mL, 5 mL 6 x 5 mL & 24 x 5 mL

AffiASSAY® Cell Counting Kit 8 (WST-8 / CCK-8)

The AffiASSAY® Cell Counting Kit 8 (WST-8 / CCK-8) is a reliable and easy-to-use assay designed for the rapid and accurate quantification of cell viability and proliferation. This kit utilizes a colorimetric method based on the reduction of a water-soluble tetrazolium salt (WST-8) by cellular dehydrogenases in viable cells to produce an orange-colored formazan dye. The intensity of the dye is directly proportional to the number of living cells in the sample, providing a sensitive measure of cell viability and proliferation.

Key Features

  • Rapid and sensitive: Provides accurate results within a short incubation period.
    Wide applicability: Compatible with a variety of cell types, including adherent and suspension cells.
  • High throughput: Suitable for high-throughput screening applications.
  • Versatile: Can be used for cell proliferation assays, cytotoxicity assays, and drug screening assays.
  • Stable signal: Offers stable and reproducible results over a wide range of cell densities and experimental conditions.

Assay Principle

The AffiASSAY® CCK-8 kit uses the water-soluble tetrazolium salt WST-8, which is reduced by cellular dehydrogenases in viable cells to form a water-soluble formazan dye. The intensity of the orange-colored formazan dye produced is directly proportional to the number of living cells in the sample. The absorbance of the formazan dye can be measured spectrophotometrically at a wavelength of 450 nm using a microplate reader.  


  • Cell viability assays: Determine the number of viable cells in culture.
  • Cell proliferation assays: Assess the rate of cell proliferation and growth.
  • Cytotoxicity assays: Evaluate the cytotoxic effects of compounds or treatments on cells.
  • Drug screening assays: Screen potential drug candidates for their effects on cell viability and proliferation. 

Kit Contents

  • CCK-8 reagent: WST-8 tetrazolium salt solution
  • User manual: Detailed instructions for assay setup and data analysis


  • For research use only. Not for diagnostic or therapeutic purposes.
  • Handle all reagents with care and follow the instructions provided in the user manual.
  • Avoid exposure of reagents to light during storage and assay procedures.
  • Use appropriate safety precautions when working with cell cultures and laboratory chemicals.

AffiASSAY® Cell Counting Kit 8 (WST-8 / CCK-8) Protocol

[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® Cell Counting Kit (CCK-8)
  • Cell culture medium
  • Cells of interest
  • 96-well microplate
  • Pipettes and tips
  • Microplate reader capable of measuring absorbance at 450 nm


Prepare Cell Culture

  • Culture cells of interest according to standard protocols in appropriate culture vessels until they reach the desired confluence or growth phase.

Prepare Working Solution

  • Thaw the CCK-8 reagent and allow it to equilibrate to room temperature.
  • Prepare the CCK-8 working solution by diluting the CCK-8 reagent with fresh cell culture medium at a ratio of 1:10 (e.g., 10 μL of CCK-8 reagent + 90 μL of medium per well). Prepare enough working solution for all wells to be assayed.

Seed Cells in Microplate

  • Aspirate the culture medium from the cells and wash once with phosphate-buffered saline (PBS) if necessary.
  • Add the appropriate volume of fresh cell culture medium containing cells to each well of a 96-well microplate. Adjust the cell density according to the experimental requirements.

Incubate Cells with CCK-8 Reagent

  • Incubate the microplate containing cells with the prepared CCK-8 working solution. Ensure each well is evenly covered with the solution.
  • Incubate the microplate at standard cell culture conditions (e.g., 37°C, 5% CO2) for a specified period of time (typically 1-4 hours). The optimal incubation time may vary depending on the cell type and experimental conditions.

Measure Absorbance

  • After the incubation period, gently mix the contents of each well by pipetting up and down or using a plate shaker.
  • Measure the absorbance of each well at 450 nm using a microplate reader. Ensure to use a reference wavelength if necessary according to the manufacturer's instructions.

Data Analysis

  • Record the absorbance values for each well.
  • Calculate the cell viability or proliferation using the formula provided by the manufacturer or based on the specific experimental design.
  • Analyze the data and interpret the results accordingly.

Quality Control

  • Perform appropriate controls, including background controls (no cells) and positive controls (e.g., untreated cells or cells treated with known compounds).
  • Ensure all reagents are handled and stored properly according to the manufacturer's instructions to maintain assay reliability.


  • If unexpected results are obtained, troubleshoot the assay by checking experimental conditions, reagent handling, and protocol adherence.
  • Consult the user manual or contact technical support for further assistance if needed

Dispose of Reagents

  • Dispose of all reagents and materials according to local regulations for biohazardous waste or chemical waste disposal.