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Cell Counting Kit-8

4194 Cited Publications
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Based on 4194 publication(s) in Google Scholar

Cell Counting Kit-8 (CCK-8) allows sensitive colorimetric assays for the determination of cell viability in cell proliferation and cytotoxicity assays.The 5 mL volume is defined as the base specification. All larger sizes correspond to incremental volumes of this base.

  • Storage :
    4°C, 2 years. Keep away from light for long-term storage.
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Cell Counting Kit-8 Product Image
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Description & Advantages

Cell Counting Kit-8 (CCK-8) allows sensitive colorimetric assays for the determination of cell viability in cell proliferation and cytotoxicity assays.

Protocol

1. Cell Number Determination

a. Inoculate cell suspension (100 μL/well) in a 96-well plate. Pre-incubate the plate in a humidified incubator (e.g., at 37°C, 5% CO2).

b. Add 10 μL of the CCK-8 solution to each well of the plate. Be careful not to introduce bubbles to the wells, since they interfere with the O.D. reading.

c. Incubate the plate for 1-4 hours in the incubator.

d. Measure the absorbance at 450 nm using a microplate reader. To measure the absorbance later, add 10 μL of 1% w/v SDS or 0.1 M HCl to each well, cover the plate and store it with protection from light at room temperature. No absorbance change should be observed for 24 hours.

2. Cell Proliferation and Cytotoxicity Assay

a. Seed cells in a 96-well plate at a density of 104-105 cells/well in 100 μL of culture medium with or without compounds to be tested. Culture the cells in a CO2 incubator at 37°C for 24 hours.

b. Add 10 μL of various concentrations of substances to be tested to the plate.

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

d. Add 10 μL of CCK-8 solution to each well of the plate using a repeating pipettor. Be careful not to introduce bubbles to the wells, since they interfere with the O.D. reading.

e. Incubate the plate for 1-4 hours in the incubator.

f. Before reading the plate, it is important to mix gently on an orbital shaker.

g. Measure the absorbance at 450 nm using a microplate reader. To measure the absorbance later, add 10 μL of 1% w/v SDS or 0.1 M HCl to each well, cover the plate and store it with protection from light at room temperature. No absorbance change should be observed for 24 hours.

Application

CCK-8 can be used to assess cell proliferation induced by cytokines and other stimulants, as well as to evaluate cytotoxicity caused by anticancer drugs or other toxic agents. It is also applicable for detecting cell growth inhibition induced by various compounds.

Storage

4°C, 2 years.

Keep away from light for long-term storage.

Attention

1. Since the CCK-8 assay is based on the dehydrogenase activity detection in viable cells, conditions or chemicals that affect dehydrogenase activity in viable cells may cause discrepancy between the actual viable cell number and the cell number determined using the CCK-8 assay.

2. WST-8 may react with reducing agents to generate WST-8 formazan. Please check the background O.D. if reducing agents are used in cytotoxicity assays or cell proliferation assays.

3. Be careful not to introduce bubbles to the wells, since they interfere with the O.D. reading.

4. If the sterilization of the CCK-8 solution is necessary, please filter the solution with a 0.2 μm membrane.

5. The incubation time varies by the type and number of cells in a well. Generally, leukocytes give weak coloration, thus a long incubation time (up to 4 hours) or a large number of cells (~105 cells/well) may be necessary.

6. Measure and subtract the O.D. at 600 nm or higher from that of sample if there is a high turbidity in the cell suspension.

7. This product is for R&D use only, not for drug, household, or other uses.

8. For your safety and health, please wear a lab coat and disposable gloves to operate.

Components

Cat. No. Product Name Package
HY-K0301-1 mL Cell Counting Kit-8 1 mL
HY-K0301-5 mL Cell Counting Kit-8 5 mL
HY-K0301-30 mL Cell Counting Kit-8 5 mL × 6
HY-K0301-120 mL Cell Counting Kit-8 5 mL × 24

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  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nature. 2025 Feb;638(8051):787-795. [Abstract]

    CCK-8 (10 μL/well; incubated at 37°C for 1-4 hours). Cell viability assays comparing the response of PC9 Del19 cancer cells and normal epithelial to the EGFR TransTAC (upper panels) or combination chemotherapy (lower panels). EGFR TransTAC inhibited PC9 Del19 cancer cells and spared normal cells, while the carboplatin and paclitaxel combination chemotherapy was toxic to both cancer and normal cell lines. A CCK-8 assay was used to measure cell counts of PC9 Del19 lung cancer cells and normal cells NHBE (normal bronchial epithelial cells), 16HBE (normal bronchial epithelial cells), BEAS-2B (normal bronchial epithelial cells) and MCF10A (normal breast epithelial cells).

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Adv Mater. 2025 Feb;37(8):e2413728. [Abstract]

    For the CCK-8 assay, 500 µL of CCK-8 solution was added to each well and incubated for 2 hours at 37 °C. Absorption at 450 nm in CCK-8 assay of different incubation groups after 12, 24, and 48 hours of incubation. The CCK-8 assay showed a significant increase in absorbance from 12 to 48 hours in the LM-Ag-pSBS group.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: J Adv Res. 2025 Aug:74:609-620. [Abstract]

    The cell viability of MH7A cells treated with methotrexate (1μM) for 24 h was determined by the CCK-8 method.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Signal Transduct Target Ther. 2024 Mar 8;9(1):63. [Abstract]

    CCK8 (1-2 h) assay-based detection of IC50 of gemcitabine in ICC cells.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Ann Rheum Dis. 2024 Sep 4:ard-2024-225565. [Abstract]

    CCK-8 (10 µL/well). Viability of RA-FLSs after daily treatment with phosphate buffered saline (PBS) or 400 nM negative control (NC), SAPT4 or SAPT8 for 5 days as determined by Cell Counting Kit-8 assays.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Drug Resist Updat. 2024 Mar:73:101052. [Abstract]

    CCK-8 (10 μL/well; 1-4 h). HCC cells, divided into DUSP4-vehicle and DUSP4-OE (overexpression) groups, were exposed to different concentrations of sorafenib (0, 2.5, 5, 10, 15, and 20 µM) for 24 h, followed by assessment of cell viability via the CCK8 assay.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: J Extracell Vesicles. 2024 Jul;13(7):e12473. [Abstract]

    CCK-8 (10 μL/well; 1 h). The HUVECs proliferation in each group determined by CCK‐8 assay (n = 3/group).

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Adv Funct Mater. 2024 Jul 16.

    CCK-8 (10 μL/well; 2 h). The cytotoxicity of OMV-CD47/GPC3 on Hepa1-6, HK-2, and L-02 cells.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: ACS Nano. 2024 Oct 29;18(43):29779-29793. [Abstract]

    CCK-8 (10 μL/well; 50 min). Quantitative determination of neuroprotection in the SH-SY5Y cell model for neuronal damage induced by Aβ1−42 oligomers.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nano Energy. 2024 Sep.

    CCK-8 (10 μL/well; 2 h). Cell viability of L929 cultured in the serum-supplemented MEM basic containing UPN extract or not after 1, 2 and, 3 days (n = 3).

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Animals (Basel). 2024 Jul 15;14(14):2072. [Abstract]

    Noggin (0-250 ng/mL; 24 h). CCK8 assay of the cytotoxicity of noggin on yak PASMCs.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nucleic Acids Res. 2023 Nov 27;51(21):11439-11452. [Abstract]

    Cell Counting Kit-8 (CCK-8) cytotoxicity assays on HGC-27 cells. Cells were incubated with 0-1000 nM of L-Apt12-6 for 48 h, and then were incubated with CCK-8 (10 μL/well) for 1 h to evaluate cytotoxicity.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nucleic Acids Res. 2023 Nov 27;51(21):11439-11452. [Abstract]

    Cell Counting Kit-8 (CCK-8) cytotoxicity assays on HeLa cells. Cells were incubated with 0-1000 nM of L-Apt12-6 for 48 h, and then were incubated with CCK-8 (10 μL/well) for 1 h to evaluate cytotoxicity.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nucleic Acids Res. 2023 Nov 27;51(21):11439-11452. [Abstract]

    Cell Counting Kit-8 (CCK-8) cytotoxicity assays on HEK293T cells. Cells were incubated with 0-1000 nM of L-Apt12-6 for 48 h, and then were incubated with CCK-8 (10 μL/well) for 1 h to evaluate cytotoxicity.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: J Mol Liq. 2023 Mar 2.

    CCK-8 (10 μM; 24 h) is used to detect the survival rate of L929 cells after treatment with different concentrations of DESs.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: J Nanobiotechnology. 2022 Jul 14;20(1):324. [Abstract]

    CCK-8 (10 μM; 1-2 h) assay was used to assess the effect of huc-MSCs-derived exosomes (1, 10, 50, 100 µg/ml) on BV2 cell viability, and ns indicates no significance.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2018 Feb 7;9(2):177. [Abstract]

    CCK8 assay. Proliferation of CAR T cells during expansion in response to repetitive stimulation with EGFR antigen.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Oncol Lett. 2018 Apr;15(4):4129-4136. [Abstract]

    The effect of TRPM8‑knockdown on PC3 cells proliferation was measured by CCK‑8 (4 h) assay. Growth of siTRPM8 cells was significantly suppressed at day 2 compared with parental and siCON cells.

  • Experimental Validation Results for Cell Counting Kit-8

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Oncol Lett. 2018 Apr;15(4):4129-4136. [Abstract]

    The effect of TRPM8‑knockdown on LNCaP cells proliferation was measured by CCK‑8 (4 h) assay. Growth of siTRPM8 cells was significantly suppressed at day 2 compared with parental and siCON cells.

Assay Types
  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nature. 2025 Feb;638(8051):787-795. [Abstract]

    CCK-8 (10 μL/well; incubated at 37°C for 1-4 hours). Cell viability assays comparing the response of PC9 Del19 cancer cells and normal epithelial to the EGFR TransTAC (upper panels) or combination chemotherapy (lower panels). EGFR TransTAC inhibited PC9 Del19 cancer cells and spared normal cells, while the carboplatin and paclitaxel combination chemotherapy was toxic to both cancer and normal cell lines. A CCK-8 assay was used to measure cell counts of PC9 Del19 lung cancer cells and normal cells NHBE (normal bronchial epithelial cells), 16HBE (normal bronchial epithelial cells), BEAS-2B (normal bronchial epithelial cells) and MCF10A (normal breast epithelial cells).

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Adv Mater. 2025 Feb;37(8):e2413728. [Abstract]

    For the CCK-8 assay, 500 µL of CCK-8 solution was added to each well and incubated for 2 hours at 37 °C. Absorption at 450 nm in CCK-8 assay of different incubation groups after 12, 24, and 48 hours of incubation. The CCK-8 assay showed a significant increase in absorbance from 12 to 48 hours in the LM-Ag-pSBS group.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: J Adv Res. 2025 Aug:74:609-620. [Abstract]

    The cell viability of MH7A cells treated with methotrexate (1μM) for 24 h was determined by the CCK-8 method.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Signal Transduct Target Ther. 2024 Mar 8;9(1):63. [Abstract]

    CCK8 (1-2 h) assay-based detection of IC50 of gemcitabine in ICC cells.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Ann Rheum Dis. 2024 Sep 4:ard-2024-225565. [Abstract]

    CCK-8 (10 µL/well). Viability of RA-FLSs after daily treatment with phosphate buffered saline (PBS) or 400 nM negative control (NC), SAPT4 or SAPT8 for 5 days as determined by Cell Counting Kit-8 assays.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Drug Resist Updat. 2024 Mar:73:101052. [Abstract]

    CCK-8 (10 μL/well; 1-4 h). HCC cells, divided into DUSP4-vehicle and DUSP4-OE (overexpression) groups, were exposed to different concentrations of sorafenib (0, 2.5, 5, 10, 15, and 20 µM) for 24 h, followed by assessment of cell viability via the CCK8 assay.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: J Extracell Vesicles. 2024 Jul;13(7):e12473. [Abstract]

    CCK-8 (10 μL/well; 1 h). The HUVECs proliferation in each group determined by CCK‐8 assay (n = 3/group).

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Adv Funct Mater. 2024 Jul 16.

    CCK-8 (10 μL/well; 2 h). The cytotoxicity of OMV-CD47/GPC3 on Hepa1-6, HK-2, and L-02 cells.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: ACS Nano. 2024 Oct 29;18(43):29779-29793. [Abstract]

    CCK-8 (10 μL/well; 50 min). Quantitative determination of neuroprotection in the SH-SY5Y cell model for neuronal damage induced by Aβ1−42 oligomers.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nano Energy. 2024 Sep.

    CCK-8 (10 μL/well; 2 h). Cell viability of L929 cultured in the serum-supplemented MEM basic containing UPN extract or not after 1, 2 and, 3 days (n = 3).

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Animals (Basel). 2024 Jul 15;14(14):2072. [Abstract]

    Noggin (0-250 ng/mL; 24 h). CCK8 assay of the cytotoxicity of noggin on yak PASMCs.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nucleic Acids Res. 2023 Nov 27;51(21):11439-11452. [Abstract]

    Cell Counting Kit-8 (CCK-8) cytotoxicity assays on HGC-27 cells. Cells were incubated with 0-1000 nM of L-Apt12-6 for 48 h, and then were incubated with CCK-8 (10 μL/well) for 1 h to evaluate cytotoxicity.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nucleic Acids Res. 2023 Nov 27;51(21):11439-11452. [Abstract]

    Cell Counting Kit-8 (CCK-8) cytotoxicity assays on HeLa cells. Cells were incubated with 0-1000 nM of L-Apt12-6 for 48 h, and then were incubated with CCK-8 (10 μL/well) for 1 h to evaluate cytotoxicity.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Nucleic Acids Res. 2023 Nov 27;51(21):11439-11452. [Abstract]

    Cell Counting Kit-8 (CCK-8) cytotoxicity assays on HEK293T cells. Cells were incubated with 0-1000 nM of L-Apt12-6 for 48 h, and then were incubated with CCK-8 (10 μL/well) for 1 h to evaluate cytotoxicity.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: J Mol Liq. 2023 Mar 2.

    CCK-8 (10 μM; 24 h) is used to detect the survival rate of L929 cells after treatment with different concentrations of DESs.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: J Nanobiotechnology. 2022 Jul 14;20(1):324. [Abstract]

    CCK-8 (10 μM; 1-2 h) assay was used to assess the effect of huc-MSCs-derived exosomes (1, 10, 50, 100 µg/ml) on BV2 cell viability, and ns indicates no significance.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Cell Death Dis. 2018 Feb 7;9(2):177. [Abstract]

    CCK8 assay. Proliferation of CAR T cells during expansion in response to repetitive stimulation with EGFR antigen.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Oncol Lett. 2018 Apr;15(4):4129-4136. [Abstract]

    The effect of TRPM8‑knockdown on PC3 cells proliferation was measured by CCK‑8 (4 h) assay. Growth of siTRPM8 cells was significantly suppressed at day 2 compared with parental and siCON cells.

  • Experimental Validation Results for Cell Counting Kit-8
    1/16

    Cell Counting Kit-8 purchased from MedChemExpress. Usage Cited in: Oncol Lett. 2018 Apr;15(4):4129-4136. [Abstract]

    The effect of TRPM8‑knockdown on LNCaP cells proliferation was measured by CCK‑8 (4 h) assay. Growth of siTRPM8 cells was significantly suppressed at day 2 compared with parental and siCON cells.

  • Cell Counting Kit-8 Product Image
    1/16
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Experimental Validation Results for Cell Counting Kit-8
  • Cell Counting Kit-8 Product Image