Cell proliferation and toxicity detection using Soleibao CCK-8 assay kit

Solar BioSolarbio CA1210 CCK-8test kit(Cell proliferation and toxicity detection kit)

Cell proliferation and toxicity detection using Soleibao CCK-8 assay kit

Product Item Number:CA1210

Product Name:CCK-8test kit(Cell proliferation and toxicity detection kit)

Product Specifications100T/500T/1000T/10*500T/10*1000T

Product Introduction:

CCK-8test kit(Cell proliferation and toxicity detection kit)

The specific product parameters are subject to the parameters in the product manual received

Specifications10*1000T ; 10*500T ; 1000T ; 100T ; 500T

Storage conditions:4℃ sealed to avoid light

Cell Counting Kit-8abbreviationCCK-8Reagent kit, forMTTThe alternative method of law is based onWST(Water soluble tetrazolium salt, chemical name:)2-(2-methoxy-4-Nitrophenyl group)-3-(4-Nitrophenyl group)-5-(2,4-Disulfobenzene)-2H-The widely used rapid and highly sensitive detection kit for cell proliferation and cytotoxicity is tetrazole monosodium salt. Can be used for drug screening, cell proliferation assay, cytotoxicity assay, tumor drug sensitivity test, etc.

Instructions for Use:
1、 Create a standard curve (when measuring the specific number of cells)
1First, count the number of cells in the prepared cell suspension using a cell counting plate, and then inoculate the cells.
2Dilute the cells in equal proportions with the culture medium to form a concentration gradient. Generally, it is necessary to3-5Cell concentration gradient, per group3-6A compound hole.
3After inoculation, culture until the cells adhere to the wall, and then addCCK-8Measure after culturing the reagent for a certain period of timeODCreate a value chart with the number of cells as the horizontal axis（XAxis),ODThe value is on the vertical axis（YThe standard curve of the 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 their addition)CCK-8The subsequent cultivation time. ）

2、 Cell viability testing

1In 96 Inoculate cell suspension into the orifice plate（100µL/Hole). Place the culture plate in the incubator for pre cultivation (in37℃,5% CO2Under the given conditions.

2Add to each hole10µL CCK-8Solution (be careful not to generate bubbles in the pores, they will affect)ODThe reading of the value.

3Incubate the culture plate in the incubator1-4Hour.

4Measure with an enzyme-linked immunosorbent assay (ELISA) reader450nmThe absorbance at the location.

5If not determined temporarilyODIf we plan to measure the value in the future, we can add it to each well10µL 0.1MofHClor1%SDS(W/V)Solution, cover the culture plate and store it at room temperature in the dark. in 24 The absorbance will not change within hours.

3、 Cell proliferation-Toxicity testing

1In 96 Configuration in orifice plate100 µLThe cell suspension. Pre cultivate the culture plate in the incubator 24 Hour (in)37 ℃，5% CO2Under the given conditions.

2Add to the culture plate10µLSubstances of different concentrations to be tested. Incubate in the incubator for an appropriate period of time (e.g.:6、12、24 or 48 Hour).

3Add to each hole10 µL CCK-8Solution (be careful not to generate bubbles in the pores, they will affect)ODThe reading of the value. If the substance to be tested has oxidizing or reducing properties, it can be addedCCK-8Previously, replace the fresh culture medium (remove the culture medium, wash the cells twice with the culture medium, and then add new culture medium) to remove the drug effect.

4Incubate the culture plate in the incubator1-4Hour.

5Measure with an enzyme-linked immunosorbent assay (ELISA) reader450 nmThe absorbance at the location.

6If not determined temporarilyODIf we plan to measure the value in the future, we can add it to each well10µL 0.1MofHClor1%SDS(W/V)Solution, cover the culture plate and store it at room temperature in the dark. in 24 The absorbance will not change within hours.

Vitality calculation:.

Cell viability (%)=[A(dosing)－A(blank)]／[ A(0dosing)－A(blank)]×100

A(Medication): with cellularCCK-8The absorbance of pores in solutions and drug solutions

A(Blank): Equipped with culture medium andCCK-8The absorbance of pores in a solution without cells

A（0 Medication: with cellsCCK-8The absorbance of pores in solutions without drug solutions

Cell viability: Cell proliferation viability or cytotoxic viability

Notes:

①Suggest making a few holes first to explore the number of cells to be inoculated and their additionCCK-8Cultivation time after reagents.

②White blood cells may need to be cultured for a longer period of time.

③When using standards 96 When using an orifice plate, the minimum inoculation amount of adherent cells should be at least1 ,000a/Kong(100 µLculture medium)The sensitivity of detecting white blood cells is relatively low, so it is recommended that the vaccination amount should not be less than2,500a/Kong( 100 µLculture medium)If you want to use 24 Orifice plate or 6 For the well plate experiment, please first calculate the corresponding inoculation amount for each well and divide it by the total volume of the culture medium in each well10%joinCCK-8Solution.

④if not450nmThe filter can be used to measure absorbance430-490 nmThe filter between them, but450nmThe detection sensitivity is the highest.

⑤The 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.

Cell proliferation and toxicity detection using Soleibao CCK-8 assay kit

Product ordering information:

CA1210 CCK-8test kit(Cell proliferation and toxicity detection kit) 100T/500T/1000T/10*500T/10*1000T