Human retinal pigment epithelium cell lines (ARPE-19) were purchased from the Academy of Sciences (Shanghai, China). Fetal bovine serum (10%, Gibco, USA) in DMEM/F12 cell medium (Hyclone, USA) with penicillin/streptomycin was used for cell culture. Cell counting kit- (CCK-) 8 and the Reactive Oxygen Species Assay Kit were purchased from Yeasen (Shanghai, China). tBHQ (30 μM) dissolved in DMSO was purchased from Topscience (Shanghai, China). Primary antibodies for Nrf2, IL-1β, IL-18, and caspase-1 were purchased from Abcam (Abcam Inc., UK). TXNIP and Trx1 were procured from Proteintech (Proteintech, USA). β-Actin and lamin B1 antibodies were purchased from CST (CST, USA); goat anti-rabbit IgG antibody (1 : 5000) from ABclonal Technology, Wuhan, China; goat anti-Mouse IgG antibody (1 : 2000) from Sigma-Aldrich. Trx1 and control siRNA were purchased from RiboBio (Guangzhou, China). C57BL/6 mice were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Beijing, China).
Cell counting kit cck 8
Manufactured by Yeasen
Sourced in China
The Cell Counting Kit (CCK-8) is a colorimetric assay used to determine the number of viable cells in a sample. It is based on the reduction of a tetrazolium salt to a water-soluble formazan dye, which can be measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Microplate reader, by Thermo Fisher Scientific (6 mentions)
High fidelity pcr enzyme 2 phanta max master mix, by Vazyme (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Luciferase detection kit, by Promega (2 mentions)
Pmd18 t 6011, by Takara Bio (2 mentions)
Taq master mix, by Vazyme (2 mentions)
Plasmid extraction kit, by Tiangen Biotech (2 mentions)
Gel extraction kit, by CWBIO (2 mentions)
Cell genome extraction kit, by Tiangen Biotech (2 mentions)
Reactive oxygen species assay kit, by Yeasen (1 mentions)
Goat anti rabbit igg antibody, by ABclonal (1 mentions)
Goat anti mouse igg antibody, by Merck Group (1 mentions)
Control sirna, by RiboBio (1 mentions)
Txnip, by Proteintech (1 mentions)
Fluostar omega, by BMG Labtech (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Lipid peroxidation kit, by Thermo Fisher Scientific (1 mentions)
Ab307601, by Abcam (1 mentions)
Ab125066, by Abcam (1 mentions)
Sybr green and reverse transcription kit, by Roche (1 mentions)
23 protocols using cell counting kit cck 8
1
Oxidative Stress and ARPE-19 Cells
2
Evaluating NPC Cell PTX Resistance
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These experiments were used to evaluate the PTX resistance and clonal proliferation ability of NPC cells by Cell Counting Kit (CCK)-8 (YEASEN Biotechnology, 40203ES60). With reference to literature [38 (link)], the half-maximal inhibitory concentration (IC50) was determined, and the relative resistance factor (RRF) was calculated by dividing the IC50 in the experimental group by the IC50 in the control group.
3
Gastric Cancer Cell Viability Assay
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5000 population/well of gastric cancer cells were seeded into 96-well plates and cell viabilities after transfection for the indicated times were detected by Cell Counting Kit (CCK-8) (Yeasen) according to the manufacturer’s instruction. OD values were the average absorbances of at least three duplicates.
4
Cell Viability Assay for Cytotoxicity
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The cells were exposed to OC or GEM treatment for 48 h 1 day after seeding in 96-well plates. Cell viability was measured using the Cell Counting Kit (CCK8) (Yeasen, China), and the absorbance was measured at 470 nm using a microplate reader (FLUOstar Omega, BMG Labtech, Germany). The IC50 values were calculated from dose–response curves using GraphPad Prism 5 (La Jolla, California, USA).
5
Assessing KGN cell proliferation
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KGN cell proliferation was assessed with a Cell Counting Kit (CCK-8, Yeasen, China). Cells were plated in a 96-well plate at a density of 2000 cells/well. On the second day, 10 µL of CCK-8 solution was added to each well. At different time points, the OD was measured using a microplate reader (Bio-Tek, China) at 450 nm.
The BeyoClickTM EdU-488 assay was performed according to the manufacturer’s protocol (Beyotime, China). Briefly, KGN cells or hGCs were cultured in 48-well plates for 48 h with 250 μM CTX and different concentrations of CEFFE. Then, proliferating cells were labeled with EdU, and the nuclei were labeled with Hoechst 33342. Finally, images of five random fields were captured by an inverted fluorescence microscope (Axio Vert.A1, Zeiss, Germany).
The BeyoClickTM EdU-488 assay was performed according to the manufacturer’s protocol (Beyotime, China). Briefly, KGN cells or hGCs were cultured in 48-well plates for 48 h with 250 μM CTX and different concentrations of CEFFE. Then, proliferating cells were labeled with EdU, and the nuclei were labeled with Hoechst 33342. Finally, images of five random fields were captured by an inverted fluorescence microscope (Axio Vert.A1, Zeiss, Germany).
6
Modulating Ferroptosis in Cells
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MCL was purchased from MedChemExpress (New Jersey, USA). Cell Counting Kit (CCK-8) was purchased from Yeasen Biotech Co., Ltd. (Shanghai, China). Ox-LDL was purchased from Yiyuan Biotechnology (Guangzhou, China). SYBR Green and Reverse Transcription kit were obtained from Roche (Basel, Switzerland). Primary antibody sources were listed: antibodies against GPX4 (ab125066) and xCT (ab307601) were purchased from Abcam (Cambridge, UK); KEAP1 (P586) and NRF2 (D1Z9C) were purchased from Cell Signaling Technology (Boston, USA), antibody against β-actin and Goat anti-rabbit antibody were obtained from Merck (Darmstadt, Germany). MDA Assay Kit, GSH assay, ROS Assay Kit, Total Superoxide Dismutase Assay Kit with NBT and LDH Cytotoxicity are obtained from Beyotime Biotechnology (Shanghai, China). Enzyme linked immunosorbent assay (ELISA) kits for TNF-α, IL-1β, IL-10 and IL-4 was obtained from Ebioscience (Wuhan, China). ML385 was obtained from MedChemExpress (New Jersey, USA). Lipid Peroxidation Kit was purchased from Invitrogen (California, USA); Iron Assay Kit was purchased from Leagene (Beijing, China). Adeno associated viral vector (AAV) with sh-NC and sh-NRF2 was purchased from HanBio Tech (Shanghai, China).
7
Antimicrobial Hydrogel Synthesis and Evaluation
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Sodium citrate dihydrate, sodium chloride (NaCl), ferric chloride hexahydrate (FeCl3·6H2O), anionic polyacrylamide (PAM, Mn = 3 000 000), urea and polyethylene glycol-2000 (PEG, Mw = 2000), polyoxymethylene and hydrogen peroxide (H2O2) were purchased from Sinopharm Chemical Reagent Co., Ltd (China). Dimethyl sulfoxide (DMSO) and 3,3′,5,5′-tetramethylbenzidine (TMB) were purchased from Aladdin Reagent Co., Ltd (Shanghai, China). Fetal bovine serum and Dulbecco’s Modified Eagle Medium were obtained from Gibco (USA). Phosphate-buffered saline (PBS) was purchased from HyClone (USA), Cell Counting Kit (CCK-8) and Calcein-AM/propidium iodide (PI) staining kit was purchased from Shanghai Yeasen Biotechnology Co., Ltd (China). Beef extract, peptone and agar were all purchased from Shanghai Yuanye Biotechnology Co., Ltd (China). Escherichiacoli (CICC 10389) and S. aureus (CICC 21600) were obtained from the China Center of Industrial Culture Collection (CICC, Wuhan, China).
8
Cytotoxicity Assay of MLB in A549 and MRC-5 Cells
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The cytotoxicity of MLB in A549 or MRC-5 cells was assessed using a Cell Counting Kit (CCK-8, Cat No. 40203; Yeasen Biotech Co. Ltd., Shanghai, China). A549 or MRC-5 cells were seeded at a density of 2 × 104 cells per well in 96-well plates. After growing for 8 h under normal growth conditions, cells were rendered quiescent by incubating in a serum-free medium for 24 h, and then treated with a normal medium containing various concentrations of MLB (0–200 μM). As the negative control, MLB was added to the medium without cells. After incubation for up to 24–48 h, 10 μL of the CCK8 solution was added to each well and incubated for ~1–2 h. The optical density at 450 nm was measured using a SYNERGY microplate reader (Biotek Winooski, Vermont, USA).
9
Antibody and Reagents Utilized for Research
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The following antibodies were used throughout this study: from ABclonal(Wuhan, China), anti-FLAG (AE063; 1:2500 for Western blot), anti-Myc (AE070; 1:2500 for Western blot), anti-β-Actin (AC026; 1:5000 for Western blot). From Abcam (Cambridge, UK), anti-hnRNP U (ab264142; 1:1000 for Western blot). From Proteintech (Wuhan, China), HRP goat anti-mouse IgG (15014; 1:5000 for Western blot), HRP goat anti-rabbite IgG (15015; 1:5000 for Western blot). anti-FLAG Magnetic Beads (HY-K0207) was purchased from MCE. 2 × Taq Master Mix (P112), High fidelity PCR enzyme- 2 × Phanta Max Master Mix (P515) were purchased from Vazyme (Nanjing, China). PMD18-T (6011) was purchased from Takara (Beijing, China). Cell Genome Extraction Kit (DP304), Plasmid Extraction Kit (DP103, DP108, DP117) were purchased from Tiangen (Beijing, China). Gel Extraction Kit (CW2302) was purchased from CWBIO (Nanjing, China). Luciferase detection kit (E6110) was purchased from Promega (Madison, USA). Cell Counting Kit (CCK-8) and TUNEL-Alexa Flour 640 apoptosis kit were purchased from Yeasen (Shanghai, China). Luciferase and nanoluc detection kit (E6110, N1110) was purchased from Promega (Madison, USA). Recombinant human IFN-alpha (11200-1), recombinant human IFN-beta (8499-IF) and recombinant human IFN-gamma (285-IF) were purchased from R&D Systems (UN).
10
Cell Viability and Colony Formation Assays
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Cells were maintained in 96‐well plates (2000 or 3000 cells/well) with 200 μL medium containing different doses of experimental drugs at 37°C for corresponding hours, following which 10 μL of Cell Counting Kit (CCK‐8; YEASEN and 40203ES80) was added to each well, and the plates were further incubated at 37°C in a humidified 5% CO2 atmosphere for 3‐4 hours. Finally, the absorbance at 450 nm was measured by a microplate reader (BioTek Synergy HT). For the colony formation assays, cells were seeded in 6‐well plates (1000 cells/well) and cultured with vehicle, 2.5 μmol/L pentamidine or 5 μmol/L pentamidine at 37°C for 48 hours. Then, the medium was replaced with fresh drug‐free medium and the cells were cultured continuously for 10 days (PC3 and DU145) or 15 days (LAPC4). Finally, the cells were fixed in 4% paraformaldehyde, stained with crystal violet and photographed.
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