C2C12 cells were obtained from ATCC (ATCC® CRL-1772™), and primary human myoblasts were obtained from Gibco. Both cell lines were cultured in Dulbecco's modified Eagle's medium (DMEM) (Gibco, USA) containing 10% fetal bovine serum (Gibco, USA) and 1% penicillin-streptomycin (Gibco, USA). Cells were seeded in the media and maintained at 37°C in a 5% CO2 cell incubator (Thermo, USA) until 70%-80% confluence. KYN concentrations of 1 μM and 10 μM were chosen based on serum concentrations found in healthy vs. pathological humans [10 (link)].
Co2 cell incubator
Manufactured by Thermo Fisher Scientific
Sourced in United States
The CO2 cell incubator is a laboratory equipment designed to provide a controlled environment for cell culture applications. Its core function is to maintain a stable temperature and carbon dioxide (CO2) level within the incubator chamber, which is essential for the growth and proliferation of various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Reverse transcription kit, by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
Atcc crl 1772, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Trypsin, by Sartorius (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Cell apoptosis and cycle kits, by BD (1 mentions)
Allegrax 15r high speed centrifuge, by Beckman Coulter (1 mentions)
Leica inverted fluorescence microscope, by Leica camera (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Transwell chamber, by Corning (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Solarbio (1 mentions)
Image lab and image pro image analysis systems, by Bio-Rad (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
Facs flow cytometer, by BD (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
18 protocols using co2 cell incubator
1
Analyzing C2C12 and Human Myoblast Response to KYN
2
Cytotoxicity and Apoptosis Assay Protocol
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Dulbecco’s Modified Eagle Medium (DMEM) was purchased from Gibco (Waltham, MA, USA). Trypsin was purchased from Biological Industries (Kibbutz Beit-Haemek, Israel). MTT was purchased from Solarbio (Beijing, China). Dimethyl sulfoxide (DMSO) was purchased from Zhiyuan Chemical Reagent Co., Ltd. (Tianjin, China). Cell apoptosis and cycle kits were purchased from BD Biosciences (San Jose, CA, USA). The trans-well chambers were purchased from Corning (Corning, NY, USA). The BCA protein assay kit was purchased from beyotime (Shanghai, China). Other reagents and solvents were of analytical grade or commercially available and used without further purification. The CO2 cell incubator was purchased from Thermo Fisher Scientific (Waltham, MA, USA). The allegraX-15R high-speed centrifuge was purchased from Beckman (Brea, CA, USA). The microplate reader was purchased from BioTek (Winooski, VE, USA). ACEA NovoCyte flow cytometry analyzer was purchased from BD Biosciences (USA). The Leica inverted fluorescence microscope was purchased from Leica (Wetzlar, Germany).
3
Multimodal Cell Analysis Protocol
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CO2 cell incubator (Thermo Fisher Scientific, Inc.), fluorescence microscope (Leica DMI 4000B/DFC425; Leica Microsystems GmbH, Wetzlar, Germany), NanoDrop ND-1000 spectrophotometer (NanoDrop Technology; Thermo Fisher Scientific, Inc., Wilmington, DE, USA), fluorescence RT-qPCR instrument (ABI 7500; Applied Biosystems, Foster City, CA, USA), microplate reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA), FACS flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA), Image Lab and Image-Pro image analysis systems (Bio-Rad Laboratories, Inc.).
4
Lentiviral Transduction of Lung Cancer Cells
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Human lung cancer cell lines A549, NCI-H1299 and SPC-A-1 were purchased from Cell resource center, Shanghai Institute of life sciences, Chinese Academy of Sciences (Shanghai, China) and EBC-1 was obtained from Mingzhou Biotechnology Co., Ltd (Ningbo, Zhejiang, China). A549 was cultured in McCoy’s 5A Media (Modified with Tricine) with 10% FBS, and NCI-H1299, SPC-A-1 and EBC-1 cell lines were grown in RPMI (w/o Hepes) Media containing 10% FBS. All cells were cultured in a CO2 cell incubator (Thermo) at 37°C. For cell infection by lentivirus, cells were seeded in a six-well plate, and 20 mL 1×108 TU/mL lentivirus were added, along with ENI.S and Polybrene for culturing. After 72 h, cell infection efficiency was valued by observing GFP fluorescence under a microscope.
5
Establishment and Metastatic Characterization of SACC Cell Lines
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SACC-83 cell line was established from a patient with SACC in the sublingual gland. SACC-LM cell line was derived from the parental SACC-83 through in vivo selection from the lung metastatic foci, and has a higher lung metastasis rate compared to SACC-83 [31 (link), 32 (link)]. Both cell lines were kindly provided by Dr. Sheng-Lin Li of Peking University School and Hospital of Stomatology, China. Cells were cultured in RPMI 1640 medium (GIBCO, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS, GIBCO, Carlsbad, CA, USA) at 37°C in 5% CO2 cell incubator (Thermo Scientific).
6
Isolation and Culture of CD133+ Cancer Cells
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CO2 cell incubator (ThermoFisher Scientific, USA); biosafety cabinet (Heal Force, BIOsafe12); inverted microscope (Motic, AE200); flow cytometer (BD, FACSCanto II); 60 mm cell culture dish (Corning, 430166); nucleic acid quantifier, mRNA reverse transcription kit, (ThermoFisher Scientific, USA); pancreatic digestion solution (Auragene Corporation), PBS (Auragene Corporation); CD133 MicroBead Kit-Tumor Tissue (MACS Miltenyi Biotec); fetal bovine serum (Gemini).
7
Cell Imaging Workflow Protocols
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We used CO2 cell incubator (Thermo Fisher Scientific, Waltham, MA, USA), Micro-camera system (version 2.0; Olympus, Tokyo, Japan), and enzyme standard instrument (Tecan Group Ltd., Männedorf, Switzerland).
8
Cell Characterization Techniques
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Li-7 cells were cultured at 37 °C, 5% CO2 cell incubator(Thermo Fisher,USA). The absorbance (Am) was measured at 490 nm using a microplate reader (Thermo Fisher, USA). The Morphology of cells was observed on a inverted phase contrast microscope (Olympus, China). The cell imaging was performed using Leica confocal microscope (Leica SP8 Corporation, Germany). Mean fluorescence intensity was was determined on the flow cytometer (BD Accuri ® C6 PLUS Bio-sciences, USA). The immunohistochemistry measurements were assayed by the bioluminescence microscope(Olympus BX43, Japan).
9
Cardiomyocyte Apoptosis and Oxidative Stress
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Rat cardiomyocytes (H9c2 cells) were purchased from the Cell Bank of the Chinese Academy of Sciences. The reagents and instruments used in this study are Trizol extraction solution (Thermo Fisher), reverse transcription kit (Thermo Fisher), Lipofectamine 3000 transfection reagent (Thermo Fisher), RIPA lysate, one-step TUNEL apoptosis assay kit (green fluorescence) (Shanghai Beyotime Biotechnology), DMEM medium (Hyclone, USA), fetal bovine serum (Gibco, USA), reactive oxygen species assay kit (Beijing Solarbio Life Sciences), PDE4D (Abcam, UK), cAMP (CST, USA), PKA (CST, USA), β-actin and HRP-labeled secondary antibody (Shanghai Beyotime Biotechnology), electrophoresis and transfer membrane reagents (Shanghai Beyotime Biotechnology), ECL luminescent solution (Millipore, USA), fluorescence microscope (Thermo Fisher), transfer electrophoresis tank (Bio-Rad, USA), and CO2 cell incubator (Thermo Fisher).
10
Cell Viability Assay of Hepatocarcinoma
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Hepatocarcinoma cells were isolated and seeded in 96-well plates (Corning, #353916) with 2000 cells per well. In the blank group, the medium containing FBS was added. The cells were cultured in a CO2 cell incubator (Thermo Fisher Scientific) for 24, 48, 72, and 96 h. Before testing, each well was changed into working solution (medium 100 μL, CCK-8 (Solarbio, #CA1210) solution 10 μL) under a dark condition. After incubation at 37°C for 1.5 h, the absorbance value at 450 nm (OD450) was measured with a microplate reader (Thermo Fisher Scientific, Multiskan FC).
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