Cell Counting Kit-8（EZB-CK8）

Introduction

The EZBioscience^® Cell Counting Kit-8(CCK-8) is a highly sensitive, non-radioactive colorimetric detection kit, which is based on WST-8 and used to determine the number of living cells in cell proliferation or toxicity experiments. In the presence of 1-methoxy PMS, WST-8 can be reduced to highly water-soluble orange-yellow formazan by intracellular dehydrogenase. The amount of the formazan generated is proportional to the number of living cells. Therefore, the number of living cells can be measured by measuring the absorbance of the formazan at 450 nm. CCK-8 Reagent of this kit is a ready to use reagent, which can be directly added to the cell samples, and can be detected after incubation for a certain period of time without pre-preparing various components.

Components

Note: Use 10 µl per reaction

Storage

The kit is recommended to be stored at 2 ~ 8°C and protected from light.

Precautions

1. In the first experiment, it is recommended to find the optimal number of cells plated and the optimal incubation time of CCK-8 Reagent (normal incubation time is 1 - 4 h).

2. Avoid bubbles in the experiment. When adding CCK-8 Reagent, it is recommended to add it close to the culture plate wall instead of inserting it into the culture medium.

3. The detection of this kit depends on the reaction catalyzed by dehydrogenase, so reducing substances and oxidizing substances will interfere with the detection. Please remove the interfering substances before using CCK-8 Reagent for detection.

4. When conducting drug inhibition experiments, if the drug contains metals (PbCl₂, FeCl₂, CuSO₄, etc.), it will affect the reaction of CCK-8 Reagent, and ultimately lead to a decrease in the sensitivity of detection.

5. If you want to determine the specific number of cells, it is recommended to make a standard curve first.

Representative Results

Figure 1. There is a linear relationship between the absorbance and the number of cells. Plate 293T cells (100 ul/well) in a 96-well microplate, place the plate in an incubator for 4 hours. Then add 10 ul CCK-8 Reagent to each well, incubate the plate in an incubator for 1 h, and measure the absorbance at 450 nm with a microplate reader.

For detailed information, please check the product manual.