Cell counting kit-8 (CCK-8) was purchased from Sigma-Aldrich. BCA protein quantitative kit was purchased from Boster (Wuhan, China). Mouse Nrf2, HO-1, SMAC, Cyt-C, Lamin-B, and β-actin monoclonal antibodies were purchased from Abcam. HRP-labeled goat anti-mouse IgG antibody and goat anti-rabbit IgG antibody were purchased from Santa Cruz. Commercial kit for the determination of lactate dehydrogenase (LDH) cytotoxicity was purchased from Thermo Fisher Scientific. Western blot electrophoresis and exposure system were purchased from Bio-Rad (CA, USA) and the automatic microplate reader was purchased from Yongchuang Medical Instruments (Shanghai, China). The other facilities used in this study included cell culture plate (Corning, USA), ultra-clean workbench (Yatai Kelong Instrument, Beijing, China), centrifuge (Shanghai Lu Xiangyi Centrifuge Instrument, China), cell counting plate (Germany), and carbon dioxide incubator (USA).
Cell culture plate
Manufactured by Corning
Sourced in United States, China, Canada
Cell culture plates are flat, shallow dishes made of plastic or glass that are commonly used in laboratories for growing and maintaining cells in a controlled environment. These plates provide a surface area for cells to adhere and proliferate, enabling researchers to conduct various experiments and studies related to cell biology, tissue engineering, and drug development.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (18 mentions)
Streptomycin, by Thermo Fisher Scientific (7 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Matrigel, by Corning (3 mentions)
Trypsin, by Avantor (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Tryple express, by Thermo Fisher Scientific (2 mentions)
Gelatin, by Merck Group (2 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (2 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Transwell plate, by Corning (2 mentions)
Cetylpyridinium bromide, by Merck Group (2 mentions)
Sodium glycerophosphate, by Merck Group (2 mentions)
Penicillin streptavidin, by Thermo Fisher Scientific (2 mentions)
E8 medium, by Cellapy (2 mentions)
Alp quantitative detection kit, by CWBIO (2 mentions)
77 protocols using cell culture plate
1
Cell Viability and Protein Analysis
2
Alveolar Epithelial Cell Inflammation Assay
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Human alveolar epithelial A549 cells was obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). AEE (purity 99.5%) was prepared in Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS (Lanzhou, China). LPS from Escherichia coli 055: B5 was obtained from Solarbio (Beijing, China). Ham’s F-12K medium, 0.05% Trypsin-EDTA, and fetal bovine serum were from Gibco (Grand Island, NY, USA). The enzyme-linked immunosorbent assay (ELISA) kits for determinations of IL-6, IL-1β, TNF-α, GSH, GPx, CAT, SOD, T-AOC, LDH, CRP, MPO, MDA and MIF were produced by Shanghai Mlbio (Shanghai, China). Cell counting kit-8 was purchased from Beyotime (Shanghai, China). BCA protein assay kit was supplied by Solarbio (Beijing, China). Cell culture plate was purchased from Corning (Beijing, China). Carboxymethylcellulose sodium (CMC-Na) was supplied by Tianjin Chemical Reagent Company (Tianjin, China). The other reagents with analytical grade were purchased from Sinopharm group (Shanghai, China).
3
Fenofibrate Inhibits VEGF-C/VEGFR-3 Signaling
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Dulbecco's modified Eagle's medium (DMEM) containing L-glutamine, fetal bovine serum (FBS), penicillin-streptomycin and 0.25% pancreatin-ethylenediaminetetraacetic acid (EDTA) were all purchased from Gibco/Invitrogen, Grand Island, NY, USA. ECA medium was obtained from ScienCell (San Diego, CA, USA); the cell culture plate was from Corning Inc., Corning, NY, USA; the 0.22 µm disposable filter was obtained from Millipore (Billerica, MA, USA); MTT reagent and cobalt(II) chloride (CoCl2) were both from Aladdin Reagent (Shanghai) Co., Ltd., Shanghai, China; the superoxide anion probe was from Beyotime, Shanghai, China; fenofibrate was obtained from Sigma Chemical Co., St. Louis, MO, USA; Matrigel was purchased from BD Biosciences, Bedford, MA, USA; anti-VEGFR-3 antibody (20712-1-AP) was from Proteintech, Chicago, IL, USA; anti-VEGFC antibody (sc-9047) was from Santa Cruz Biotechnology, Inc., Dallas, TX, USA; anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (P30008) and IgG-HRP (M21002) were from Abmart (Shanghai, China); PVDF membranes were purchased from Millipore; skim milk was purchased from BD Biosciences; HRP-substrate coloring solution was obtained from Millipore; the VEGFC enzyme-linked immunosorbent assay (ELISA) kit and the VEGFR-3 ELISA kit were from USCN Life Science, Inc., Wuhan, China.
4
RAW264.7 Cell Culture Protocol
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RAW264.7 cells were purchased from the American Type Culture Collection. The culture reagents included Dulbecco's modified Eagle medium (DMEM; Gibco, C11995500CP), Roswell Park Memorial Institute (RPMI) 1640 (Gibco, C11875500BT), fetal bovine serum (Bio IND, 04-002-1A), antimycotic (Lifetechnologies, 15240-112), phosphate-buffered saline (PBS), pH 7.4 (Gibco, 10010-023), trypsin-EDTA (0.05%; Lifetechnologies, 25300-054), bovine serum albumin (Lifetechnologies, 15561012), and CellTiter-Glo (CTG; PROMEGA, G7572). Consumables and instruments included a cell culture plate (Corning), cell culture flask (Corning), microplate tester (BioTek, HM-1), and conventional instruments, such as a CO2 incubator (Thermo 3111) and biosafety cabinet (Heal Force, HFSAFe-1200LC).
5
Intestinal Organoid Propagation Protocol
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Organoids were propagated by dissociating crypt-derived organoids in TryplE Express (Invitrogen) for 3 minutes at 37°C. After this time, the TryplE Express was quenched by adding 1–2 times that amount of Advanced DMEM/F12 (Gibco). The pellet containing the dissociated intestinal single cells after centrifugation in a microcentrifuge (0540390; Thermo Fisher) at 300 r.c.f. for 5 minutes was resuspended in Matrigel (356231 Growth Factor Reduced; Corning) and embedded onto a flat-bottom, 24-well cell culture plate (3526; Corning) by forming 20-μL droplets of Matrigel, creating at least 3 technical replicates for each condition. The embedded Matrigel droplets were immediately placed inside a fully humidified incubator containing 5% CO2, which was maintained at 37°C for 5 minutes to solidify the Matrigel droplets. Once the Matrigel was solidified, 600 μL supplemented Advanced DMEM/F12 cell medium described earlier was added to each well. The media was changed every 2 days for each well and the plate was maintained in a 37°C incubator.
6
Porous Gelatin-Chondroitin-Hyaluronic Scaffold
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Gelatin (5% (w/v); Sigma, San Jose, CA, USA) was dissolved in 10 ml distilled water and slowly added to chondroitin-6-sulfate (0.05% (w/v); Sigma) and hyaluronic acid (0.1% (w/v); Sigma) to form a suspension solution. The solution was stirred for 60 minutes at room temperature with a magnetic stirrer, and then cross-linker solution (0.5% (w/v) 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 0.25% (w/v) N-hydroxysuccinimide; Sigma) was added dropwise to the suspension solution and stirred for 15 minutes. The mixture was injected into wells of a cell culture plate (2 cm diameter, 1.8 cm height; Corning-costar, NY, USA). Plates were agitated horizontally to enable even cell distribution. Constructs were then frozen at -80°C for 2 hours and lyophilized with a freeze dryer (CHRIST, Vaihingen, Germany) for 48 hours. Gel-C6S-HA porous sponge-like scaffolds, with a thickness of 2 mm, were obtained. Scaffolds were soaked in 75% (v/v) ethanol followed by phosphate-buffered saline (PBS), each for 48 hours, and were dried with sterile gauze for further use. The macroscopic appearance of the Gel-C6S-HA scaffold was photographed using a digital camera (Sony, Tokyo, Japan).
7
Murine Splenocyte Cytokine Assay
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On day 28 after immunization, mice were sacrificed after anesthesia with ether and spleen lymphocytes were aseptically isolated from mice. Cells were resuspended in RPMI-1640 containing 10% FBS (v/v) and 1% penicillin–streptomycin solution (v/v). The cells added to a 96-well cell culture plate (Corning, 106 cells/well, 100 μl), 1 μl Concanavalin A (Con A) (Sigma, 1 mg/ml), and type-O FMD VLPs (1 mg/ml) were added to the wells as positive control and a blank control group containing only culture medium but not cells was established. After 72 h of culture, the culture supernatants before and after the stimulation of lymphocytes in each group were analyzed for IFN-γ content. The OD at 490 nm was detected, and finally, the stimulation index (SI) was calculated as follows:
8
Gelatin-Chitosan Hydrogel Fabrication
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Gelatin (type A, from porcine skin) and chitosan (low molecular weight) was purchased from Sigma-Aldrich (Shanghai, China). Methacrylic anhydride was purchased from Aladdin (Shanghai, China). 3,4-Dihydroxyhydrocinnamic acid, N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), ethanol (AR, 99.7%), DL-dithiothreitol (DTT) and zinc chloride (ZnCl2) were purchased from Macklin (Shanghai, China). CCK-8 proliferation assay kit was purchased from Beyotime (Shanghai, China). Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium (DMEM/F-12), induction media (osteogenic, chondrogenic and adipogenic) and fetal bovine serum (FBS) were ordered from Invitrogen. Cell culture plate was ordered from Corning.
9
Cell Viability Assay with GA Treatment
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First, 100 μl of cell suspension (50,000 cells/ml) was added to each well in a 96-well Corning™ cell culture plate. The plate was placed in an incubator at 37°C with 5% CO2 for 24 h culture. The plates were divided into six groups at different concentrations of GA including 0, 0.5, 1.0, 1.5, 2.0, and 3.0 μmol/L, and six complementary holes were set per group. A zero hole of the cell fluid was set only in the cytometry, the blank control hole that does not add drugs. The culture plates were then incubated at 37°C, and the results were observed after 24 h. A total of 10 µl CCK-8 reagent (Beyotime Biotechnology, Shanghai, China) was added to each well for additional 1 h, and the cell viability was measured using the CCK-8 assay. The absorbance at 450 nm was measured using a multidetection microplate reader (BioTek, United States), and the relative cell viability was calculated.
10
Enhancing Islet Transplantation with PD-L1 Silencing
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MSCs (1.5 × 105) were cultured on a cell culture plate (60 mm by 15 mm; Corning, Oneonta, NY, USA) for 24 hours before transfection. Next day, MSCs were incubated with 2 ml of MEM-α medium containing 50 nM PD-L1 siRNA (No60533-1; Bioneer, Daejeon, Republic of Korea) and Lipofectamine RNAiMAX Transfection reagent (Thermo Fisher Scientific, Waltham, MA, USA) for 6 hours at 37°C. Cells were then rinsed, trypsinized, and used to fabricate hybrid spheroids with RAP-MPs. To increase durations of gene silencing, hybrid spheroids were incubated with the transfection complex for 6 hours before being transplanted with islets. Transfection efficiencies and gene silencing durations were determined by qRT-PCR.
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