Intended Use
Cell Counting Kit-8 (CCK-8) allows sensitive colorimetric assays for the determination of cell viability in cell proliferation and cytotoxicity assays.
General Description
CCK solution is a one-bottle solution; no pre-mixing of components is required. CCK, being nonradioactive, allows sensitive colorimetric assays for the determination of the number of viable cells in cell proliferation and cytotoxicity assays. WSTS* is reduced by dehydrogenases in cells to give an orange colored product (formazan), which is soluble in tissue culture medium. The amount of formazan dye generated by dehydrogenases in cells is directly proportional to the number of living cells.
Comparison between CCK method and MTT method:
Protocol:
1. Usually in the 96 hole plate, cell proliferation and cytotoxicity test are inoculated with cell suspension (5000cells and 2000 cells /100 uL/ hole, the specific number of the cells per hole should be decided according to the size and speed of the cell proliferation speed and other factors). The system is filled with a corresponding amount of cell culture medium, drug and CCK-8 solution, and the holes without cells are regarded as a blank control.
2. The cells adhered to the wall for 24 hours and are given a specific dose of 0-10 mg of stimulation in accordance with the experimental requirements.
3. The drugs are treated for 2~4 days and then add 10 uL of CCK solution at each hole (be careful not to generate bubbles in the pores, which will affect OD value)
4. Incubate the incubator for 1~4 hours, then use the microplate reader to determine the absorbance at 450nm.
5. If you do not detect OD value
temporarily, you may add 10 μL 0.1M HCL solution or 1% w/v SDS
solution, cover it with culture plate and keep it from light at room
temperature. Absorbance does not change in 24 hours.
6. The absorbance of phenol red can be eliminated by blank hole so that the result will not be affected.
Announcements:
1. When using 96 hole plate to detect , if the cell culture time is longer,we must pay attention to evaporation problems. On the one hand, a circle surrounding 96 hole plate is more likely to evaporate, you can take it out, adding the same amount of PBS, water or liquid culture instead; On the other hand, the 96 hole plate can be placed near the incubator water to reduce evaporation.
2. The detection of this kit depends on the reaction catalyzed by dehydrogenase, so reducing agents (such as some antioxidants) will interfere with the detection.
3. If there are many reducing agents in the detection system, we should try to remove them.
4. Before using Microplate Reader, you should make sure there is no bubble in each hole, otherwise it will interfere with the determination.
5. This product is limited to scientific research of professionals and should not be used for clinical diagnosis and treatment or used as food and medicine, it should not be deposited in ordinary houses.
6. For your safety and health, please wear lab-gown and disposable gloves.
Vitality calculation:
Cell viability * (%) =[A (dosing) -A (blank)]/[A (0 dosing) -A (blank) * 100%
A (dosing): Absorbance of cells, CCK solutions, and drug solutions
A (blank): Absorbance of the culture medium and CCK solution without cells
A (0 dosing): Absorbance of cells, CCK solutions without drug solutions
* Cell viability: Cell proliferation activity or cytotoxicity activity
Storage conditions:
4 degrees centigrades for one year; -20 degrees centigrades, protected from light for two years.
Documentation Download
2020 Aug.
Blockage of AMPK-ULK1 pathway mediated autophagy promotes cell apoptosis to increase doxorubicin sensitivity in breast cancer (BC) cells: an in vitro study
Cell Counting Kit-8 purchased from AbMole
Life Sci. 2021 Jan 15;265:118853.
Circadian clock gene BMAL1 reduces urinary calcium oxalate stones formation by regulating NRF2/HO-1 pathway
Cell Counting Kit-8 purchased from AbMole
FEBS Open Bio. 2020 Dec 19.
Nicotinamide mononucleotide (NMN) protects bEnd.3 cells against H 2 O 2 -induced damage via NAMPT and the NF-κB p65 signalling pathway
Cell Counting Kit-8 purchased from AbMole
2020 Dec.
A Novel Circular RNA hsa_circRNA_103809/miR-377-3p/GOT1 Pathway Regulates Cisplatin-Resistance in Non-Small Cell Lung Cancer (NSCLC)
Cell Counting Kit-8 purchased from AbMole
2020 Aug.
Blockage of AMPK-ULK1 pathway mediated autophagy promotes cell apoptosis to increase doxorubicin sensitivity in breast cancer (BC) cells: an in vitro study
Cell Counting Kit-8 purchased from AbMole
Onco Targets Ther. 2020 Sep 28;13:9543-9558.
Targeted Inhibition of P4HB Promotes Cell Sensitivity to Gemcitabine in Urothelial Carcinoma of the Bladder
Cell Counting Kit-8 purchased from AbMole
Life Sci. 2020 Oct 15;259:118180.
Upregulation of miR-133a-3p enhances Bufothionine-induced gastric cancer cell death by modulating IGF1R/PI3K/Akt signal pathway mediated ER stress
Cell Counting Kit-8 purchased from AbMole
J Appl Microbiol. 2020 Sep 29.
Enhanced in vitro antimicrobial activity of amphotericin B with berberine against dual-species biofilms of Candida albicans and Staphylococcus aureus
Cell Counting Kit-8 purchased from AbMole
Exp Physiol. 2020 Sep 22.
Involvement of the lncRNA AFAP1-AS1/microRNA-195/E2F3 axis in proliferation and migration of enteric neural crest stem cells of Hirschsprung's disease
Cell Counting Kit-8 purchased from AbMole
Cell Death Dis. 2020 Aug 20;11(8):664.
Long noncoding RNA TTN-AS1 facilitates tumorigenesis and metastasis by maintaining TTN expression in skin cutaneous melanoma
Cell Counting Kit-8 purchased from AbMole
Biochem Biophys Res Commun. 2020 Sep 10;530(1):47-53.
LncRNA Linc-PINT inhibits miR-523-3p to hamper retinoblastoma progression by upregulating Dickkopf-1 (DKK1)
Cell Counting Kit-8 purchased from AbMole
2020 Jun.
The Role of Transcription Factor Ap1 in The Activation of Nrf2/ARE Pathway Through TET1 in Diabetic Nephropathy
Cell Counting Kit-8 purchased from AbMole
Aging (Albany NY). 2020 Jun 9;12(11):11025-11041.
LncRNA ADAMTS9-AS2 inhibits gastric cancer (GC) development and sensitizes chemoresistant GC cells to cisplatin by regulating miR-223-3p/NLRP3 axis
Cell Counting Kit-8 purchased from AbMole
IUBMB Life. 2020 Jun 27.
Effects of genetic variations in Acads gene on the risk of chronic obstructive pulmonary disease
Cell Counting Kit-8 purchased from AbMole
World J Gastroenterol. 2020 Jun 7;26(21):2810-2820.
Gan Shen Fu Fang ameliorates liver fibrosis in vitro and in vivo by inhibiting the inflammatory response and extracellular signal-regulated kinase phosphorylation
Cell Counting Kit-8 purchased from AbMole
Open Life Sciences. 2020 Jun 30;15(1):409-417.
Buformin suppresses osteosarcoma via targeting AMPK signaling pathway
Cell Counting Kit-8 purchased from AbMole
Anat Rec (Hoboken). 2020 Jan 21.
Resibufogenin inhibited colorectal cancer cell growth and tumorigenesis through triggering ferroptosis and ROS production mediated by GPX4 inactivation
Cell Counting Kit-8 purchased from AbMole
Sci Adv. 2020 Jul 10;6(28):eaba7260.
A basal-bolus insulin regimen integrated microneedle patch for intraday postprandial glucose control
Cell Counting Kit-8 purchased from AbMole
Exp Biol Med (Maywood). 2020 Jun;245(11):925-932.
Circular RNA circ-CDYL sponges miR-1180 to elevate yes-associated protein in multiple myeloma
Cell Counting Kit-8 purchased from AbMole
Aging (Albany NY). 2020 May 4;12(9):8137-8150.
Overexpression of METTL3 attenuates high-glucose induced RPE cell pyroptosis by regulating miR-25-3p/PTEN/Akt signaling cascade through DGCR8
Cell Counting Kit-8 purchased from AbMole
Aging (Albany NY). 2020 Jun 9;12(11):11025-11041.
LncRNA ADAMTS9-AS2 inhibits gastric cancer (GC) development and sensitizes chemoresistant GC cells to cisplatin by regulating miR-223-3p/NLRP3 axis
Cell Counting Kit-8 purchased from AbMole
Cell Cycle. 2020 Aug;19(15):1857-1868.
Hsa_circ_0009910: oncogenic circular RNA targets microRNA-145 in ovarian cancer cells
Cell Counting Kit-8 purchased from AbMole
J Exp Clin Cancer Res. 2020 May 24;39(1):93.
TFEB-NF-κB Inflammatory Signaling Axis: A Novel Therapeutic Pathway of Dihydrotanshinone I in Doxorubicin-Induced Cardiotoxicity
Cell Counting Kit-8 purchased from AbMole
J Clin Lab Anal. 2020 Feb 28;e23249.
Retraction: Hsa_circ_0003645 Shows an Oncogenic Role by Sponging microRNA-1299 in Hepatocellular Carcinoma Cells
Cell Counting Kit-8 purchased from AbMole
Biomed Pharmacother. 2020 May 25;128:110244.
Emodin Succinyl Ester Inhibits Malignant Proliferation and Migration of Hepatocellular Carcinoma by Suppressing the Interaction of AR and EZH2
Cell Counting Kit-8 purchased from AbMole
J Cell Mol Med. 2020 Mar;24(5):2901-2916.
Verbascoside Inhibits Progression of Glioblastoma Cells by Promoting Let-7g-5p and Down-Regulating HMGA2 via Wnt/beta-catenin Signalling Blockade
Cell Counting Kit-8 purchased from AbMole
Vet Res. 2020 Apr 10;51(1):53.
Newcastle Disease Virus RNA-induced IL-1β Expression via the NLRP3/caspase-1 Inflammasome
Cell Counting Kit-8 purchased from AbMole
Saudi J Gastroenterol. 2020 May-Jun;26(3):144-152.
Inhibition of ALAS1 Activity Exerts Anti-Tumour Effects on Colorectal Cancer in vitro
Cell Counting Kit-8 purchased from AbMole
Naunyn Schmiedebergs Arch Pharmacol. 2020 Feb 8.
Knock-down of LncRNA-XIST Induced Glioma Cell Death and Inhibited Tumorigenesis by Regulating miR-137/SLC1A5 Axis-Mediated ROS Production
Cell Counting Kit-8 purchased from AbMole
Eur J Pharmacol. 2020 May 26;173212.
Pinocembrin Alleviates Glucocorticoid-Induced Apoptosis by Activating Autophagy via Suppressing the PI3K/Akt/mTOR Pathway in Osteocytes
Cell Counting Kit-8 purchased from AbMole
Colcom. 2020 Jul;Volume 37, 100268.
Antibacterial coatings based on microgels containing quaternary ammonium ions: Modification with polymeric sugars for improved cytocompatibility
Cell Counting Kit-8 purchased from AbMole
Chemosphere. 2020 Aug;253:126707.
Bisphenol A and Its Analogues Bisphenol S, Bisphenol F and Bisphenol AF Induce Oxidative Stress and Biomacromolecular Damage in Human Granulosa KGN Cells.
Cell Counting Kit-8 purchased from AbMole
Cell Cycle. 2020 May;19(10):1067-1076.
Hypoxia-induced Apoptosis of Cardiomyocytes Is Restricted by Ginkgolide B-downregulated microRNA-29.
Cell Counting Kit-8 purchased from AbMole
ACS Appl Mater Interfaces. 2020 Apr 29;12(17):19268-19276.
Universal Antibacterial Surfaces Fabricated From Quaternary Ammonium Salt-Based PNIPAM Microgels.
Cell Counting Kit-8 purchased from AbMole
Onco Targets Ther. 2020 May 4;13:3729-3737.
Circular RNA circTUBGCP3 Is Up-Regulated and Promotes Cell Proliferation, Migration and Survivability via Sponge mir-30b in Osteosarcoma.
Cell Counting Kit-8 purchased from AbMole
Biomed Pharmacother. 2020 May;125:109924.
MiR-130a Alleviated High-Glucose Induced Retinal Pigment Epithelium (RPE) Death by Modulating TNF-α/SOD1/ROS Cascade Mediated Pyroptosis.
Cell Counting Kit-8 purchased from AbMole
Carbohydr Polym. 2020 Jan 1;227:115314.
Structural characterisation and immunomodulatory activity of polysaccharides from white asparagus skin.
Cell Counting Kit-8 purchased from AbMole
Biofactors. 2020 Jan 8.
Propofol exerts neuroprotective functions by down-regulating microRNA-19a in glutamic acid-induced PC12 cells.
Cell Counting Kit-8 purchased from AbMole
Anat Rec (Hoboken). 2020 Jan 21.
Resibufogenin inhibited colorectal cancer cell growth and tumorigenesis through triggering ferroptosis and ROS production mediated by GPX4 inactivation.
Cell Counting Kit-8 purchased from AbMole
Yonsei Med J. 2019 Sep;60(9):832-841.
miR-1301/TRIAP1 Axis Participates in Epirubicin-Mediated Anti-Proliferation and Pro-Apoptosis in Osteosarcoma.
Cell Counting Kit-8 purchased from AbMole
Oncol Lett. 2019 Nov;18(5):4613-4620.
MicroRNA-152 suppresses cisplatin resistance in A549 cells.
Cell Counting Kit-8 purchased from AbMole
Med Sci Monit. 2019 Aug 11;25:5977-5985.
Expression of circZNF609 is Down-Regulated in Colorectal Cancer Tissue and Promotes Apoptosis in Colorectal Cancer Cells by Upregulating p53.
Cell Counting Kit-8 purchased from AbMole
J Hazard Mater. 2020 Apr 15;388:121802.
Thiol-functionalized inactivated yeast embedded in agar aerogel for highly efficient adsorption of patulin in apple juice.
Cell Counting Kit-8 purchased from AbMole
J Clin Neurosci. 2019 Jun;64:234-241.
Potential contribution of microRNA-125b targeting p38MAPK to relieving intermittent hypoxia-induced dementia of rat models.
Cell Counting Kit-8 purchased from AbMole
Biochem Biophys Res Commun. 2019 Apr 30;512(2):218-223.
Versatile Pt NCs-based chemotherapeutic agents significantly induce the apoptosis of cisplatin-resistant non-small cell lung cancer.
Cell Counting Kit-8 purchased from AbMole
International Journal of Oncology. 2019 Mar;929-941.
lncRNA H19 promotes viability and epithelial-mesenchymal transition of lung adenocarcinoma cells by targeting miR-29b-3p and modifying STAT3.
Cell Counting Kit-8 purchased from AbMole
Int J Pharm. 2018 Dec 13;556:338-348.
Self-implanted tiny needles as alternative to traditional parenteral administrations for controlled transdermal drug delivery.
Cell Counting Kit-8 purchased from AbMole
Med Sci Monit. 2018 Oct 28;24:7710-7718.
Knockdown of MON1B Exerts Anti-Tumor Effects in Colon Cancer In Vitro.
Cell Counting Kit-8 purchased from AbMole
Sci Rep. 2018 Mar 1;8(1):3891.
Bufalin suppresses hepatocarcinogenesis by targeting β-catenin/TCF signaling via cell cycle-related kinase
Cell Counting Kit-8 purchased from AbMole
Cancer Biol Ther. 2018 Feb 5.
Buformin suppresses proliferation and invasion via AMPK/S6 pathway in cervical cancer and synergizes with paclitaxel
Cell Counting Kit-8 purchased from AbMole
Future Oncology 2018 Jan 10.
The overexpression of PXN promotes tumor progression and leads to radioresistance in cervical cancer
Cell Counting Kit-8 purchased from AbMole
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