CCK-8 kit

CCK8 is mainly used for cell viability proliferation and toxicity detection. To purchase this kit, you need to bring your own 96-well plate, 10 u, 100-200 ul and multi-channel pipette, a microplate reader with a 450 nm filter, and CO2 culture box.

Reagent specifications: 1ml/100T 

cell activity detection
1. Inoculate cell suspension (100/) in a 96-well plate.Place the culture plate in an incubator for pre-culture (at 37C, 5% CO2).
2. Add 10u of CCK-8 solution to each well (be careful not to generate bubbles in the wells, they will affect the OD value reading)
3. Incubate the culture plate in the incubator for 1-4 hours.
4. Use a microplate reader to measure the absorbance at 450 nm.
5. If you do not want to measure the OD value temporarily and plan to measure it later, you can add 10 ul01 M HCI solution or 1% w/v SDS solution to each well, and cover the culture plate to avoid light and store it at room temperature until the absorbance does not change within 24 hours. Changes will occur.

Cell Proliferation-Toxicity Assay
1. Prepare 100 cell suspensions in 96 plates.Pre-incubate the culture plate in the incubator for 24 hours (at 37C, 5% CO). 2. Add 10 l of different concentrations of the substance to be tested to the culture plate.
3. Incubate the culture plate in the incubator for an appropriate period of time (for example: 6, 24 or 48 hours).
4. Add 10 CCK-8 solution to each well (be careful not to generate bubbles in the wells, they will affect the OD value reading).
5. Incubate the culture plate in the incubator for 1-4 hours.
6. Use a microplate reader to measure the absorbance at 450 nm.
7. If you do not measure the OD value temporarily and plan to measure it later, you can add 10 ul 0.1 M HCI solution or 1% w/v SDS solution to each well and cover the culture. The absorbance will not change within 24 hours if the plate is stored at room temperature in the dark.Note: If the substance to be tested has oxidizing or reducing properties, the medium can be replaced with fresh culture medium before adding CCK-8 to remove the influence of the drug.Of course, if the effect of the drug is relatively small, you do not need to replace the culture medium, and you can directly deduct the blank absorption after adding the drug to the culture medium.

Preparing a standard curve
1. First count the number of cells in the prepared cell suspension using a cell counting plate, and then inoculate the cells.
2 According to the ratio (for example: 1/2 ratio), dilute the medium into a cell concentration gradient in equal proportions. Generally, 3-5 cell concentration gradients are made, with 3-6 duplicate wells in each group.
3. After inoculation, culture 2- Allow the cells to adhere to the wall for 4 hours, then add CCK-8 reagent and incubate for a certain period of time, then measure the OD value, and create a standard curve with the number of cells as the abscissa (X-axis) and the OD value as the ordinate (Y-axis).The number of cells in unknown samples can be determined based on this standard curve (the prerequisite for using this standard curve is that the experimental conditions must be consistent, so as to facilitate the determination of the number of cells inoculated and the culture time after adding CCK-8) 

Statement: The product is for scientific research use only