ZETA LIFE Cell Proliferation and Toxicity Detection CCK Kit

ZETA LIFE Cell Proliferation and Toxicity Detection CCK Kit

Price:￥615 - 2800

Brand:ZETA LIFE .

Item No.K009

Place of OriginUSA

supplier:ShanghaiLai TongBiotechnology Co., Ltd

quantity:1000

English name:cck8

shelf life:24 months

Storage conditionsStore at 4 ° C in the dark

specification:500T-3000T

ZETA LIFE Cell Proliferation and Toxicity Detection CCK Kit

CellCounting Kit, abbreviated as CCK kit, is a rapid and highly sensitive detection kit based on WST (water-soluble tetrazolium salt, chemical name: 2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2,4-disulfophenyl) -2H tetrazolium monosodium salt), widely used for cell proliferation and cytotoxicity.

CCK is an upgraded alternative product to MTT, which has significant advantages compared to MTT or other MTT similar products such as XTT. MTS

-The formazan produced by CCK reduced by dehydrogenase in mitochondria is water-soluble and does not require a specific solution for dissolution, while the formazan produced by WST, XTT, and MTS are not water-soluble, thus eliminating the need for subsequent dissolution steps.

-CCK is more stable than XTT and MTS, making the experimental results more stable.

-Compared with MTT, XTT, etc., CCK has a wider linear range and higher sensitivity.

Comparison between CCK method and other detection methods:

Operating instructions:

1、 Create a standard curve (when measuring the specific number of cells)

1. First, count the number of cells in the prepared cell suspension using a cell counting plate, and then inoculate the cells.

2. Dilute the culture medium in equal proportions (e.g. 1/2 ratio) to form a cell concentration gradient. Generally, 3-5 cell concentration gradients are required, with 3-6 replicates per group.

3. After inoculation, culture the cells for 2-4 hours to allow them to adhere to the wall, then add CCK reagent to culture for a certain period of time and measure the OD value. Create a line with the number of cells as the x-axis,

The OD value is the standard curve of the y-axis. According to this standard curve, the number of cells in an unknown sample can be determined (the premise of using this standard curve is that the experimental conditions must be consistent)

To facilitate the determination of the number of cells inoculated and the cultivation time after adding CCK.

2、 Cell viability testing

1. Inoculate cell suspension (100ul well) into a 96 well plate. Place the culture plate in the incubator for pre cultivation (at 379C, 5% CO2).

2. Add 110ul of CCK solution to each well (be careful not to generate bubbles in the well, as they can affect the OD reading).

3. Incubate the culture plate in the incubator for 14 hours.

4. Measure the absorbance at 450nm using an enzyme-linked immunosorbent assay (ELISA) reader.

5. If the OD value is not measured temporarily and plans to be measured in the future, 10ul of 0.1M HCL solution or 1% wIv SDS solution can be added to each well, and the culture can be covered

Store the board in the dark at room temperature. The absorbance will not change within 24 hours.

3、 Cell proliferation~toxicity detection

1. Prepare a 100u cell suspension in a 96 well plate. Pre culture the culture plate in the incubator for 24 hours at 379C and 5% CO2.

2. Add 10ul of different concentrations of the test substance to the culture plate.

3. Incubate the culture plate in the incubator for an appropriate period of time (e.g. 6, 12, 24, or 48 hours).

4. I want to add 10ul of CCK solution to each well (be careful not to generate bubbles in the well, as they will affect the OD reading).

5. Incubate the culture plate in the incubator for 1-4 hours.

6. Measure the absorbance at 450nm using an enzyme-linked immunosorbent assay (ELISA) reader.

7. If the OD value is not measured temporarily and is planned to be measured in the future, 10ul of 0.1M HCL solution or 1% wIv SDS solution can be added to each well, and the culture plate can be covered and stored at room temperature in the dark. The absorbance will not change within 24 hours.

Attention: If the substance to be tested has oxidizing or reducing properties, fresh culture medium can be replaced before adding CCK (remove the culture medium, wash the cells twice with the culture medium, and then add new culture medium) to remove drug effects. Of course, in cases where the drug has a relatively small impact, the blank absorption after adding the drug to the culture medium can be directly deducted without changing the culture medium.

To be prepared:

①Suggest making a few wells first to explore the number of cells to be inoculated and the cultivation time after adding CCK reagent.

②If conditions permit, it is recommended to use a multi-channel pipette to reduce the difference in parallel hole reduction. join

When adding CCK reagent, it is recommended to add it diagonally against the wall of the culture plate and not insert it under the culture medium page, as it may generate bubbles and interfere with the OD value reading.

③White blood cells may need to be cultured for a longer period of time.

④When using a standard 96 well plate, the small inoculation volume of adherent cells should be at least 1000 cells per well L (100u culture medium). The sensitivity of detecting white blood cells is relatively low, therefore

The recommended vaccination dose is not less than 2500/F wells (100ul of culture medium). If you want to use a 24 well or 6-well plate for the experiment, please first calculate the corresponding inoculation amount for each well and add CCK solution at 10% of the total volume of the culture medium for each well.

⑤If there is no 450nm filter, a filter with absorbance between 430-490nm can be used, but the detection sensitivity of 450nm filter is higher.

⑥The absorbance of phenol red in the culture medium can be eliminated by subtracting the absorbance of the background in the blank well during calculation, so it will not affect the detection.

Dynamic calculation:

Cell viability * (%)=[A (dosing) - A (blank)]/[A (0 dosing) - A (blank)] x100

A (Medication): Absorbance of pores containing cells, CCK solution, and drug solution

A (blank): Absorbance of wells with culture medium and CCK solution without cells

A (0 dosing): absorbance of wells with cells, CCK solution but no drug solution

*Cell viability: Cell proliferation viability or cytotoxic viability

Storage conditions:

Store at 49C away from light, with a shelf life of one year.

Warm sound prompt: Not suitable for clinical treatment.