A549 cells were grown in PRIM-1640 (Invitrogen, USA) with 10% heat-inactivated fetal bovine serum and antibiotics (100 µg/mL of streptomycin and 100 U/mL of penicillin) at 37 °C with 5% CO2. Cells were seeded in 24-well plates and grown overnight prior to studies. Cells were washed two times with phosphate-buffered saline (PBS, pH = 7.4), incubated with dye-labeled DNA nanostructures (the finial concentration was estimated as 50 nM of labeled dyes for each structure) in cell culture medium (500 μL), containing FBS [10% (vol/vol)] or not at 37 °C for different times (2, 6,12, 24 h), followed by washing with PBS three times. Then, cells were digested with 0.25% trypsin (Gibco, USA) and resuspended in PBS (200 μL). Cells with the same procedure but without DNA nanostructures were used as controls. Samples of at least 5000 cells were analyzed in triplicate using flow cytometry (FACSArray; BD Biosciences, San Jose, CA, USA). Consistent gating based on cell size and granularity (forward and side scatter) was applied to select only fluorescence measurements from healthy cells and exclude cell debris and doublets. Gating region indicates the main population of the cells we analyzed (Supplementary Figure 19 ).
Prim 1640
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PRIM 1640 is a compact and reliable laboratory centrifuge designed for general-purpose applications. It features a maximum speed of 16,000 rpm and a maximum RCF of 25,000 x g, making it suitable for a variety of sample preparation and separation tasks. The PRIM 1640 is equipped with a brushless motor for quiet and efficient operation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Ges 1, by Shanghai Institute of Biochemistry and Cell Biology (2 mentions)
Sgc 7901, by Shanghai Institute of Biochemistry and Cell Biology (2 mentions)
Penicillin and streptomycin, by Merck Group (2 mentions)
Facsarray, by BD (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Dksfm, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Epilife, by Thermo Fisher Scientific (1 mentions)
Mycoplasma real time pcr detection kit, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by ExCell Bio (1 mentions)
Mda mb 231, by Cell Resource Center (1 mentions)
A549 ddp, by National Collection of Authenticated Cell Cultures (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
A549 ddp, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
H1299, by National Collection of Authenticated Cell Cultures (1 mentions)
Verteporfin, by Merck Group (1 mentions)
Bgc 823, by Shanghai Institute of Biochemistry and Cell Biology (1 mentions)
C11995500bt, by Thermo Fisher Scientific (1 mentions)
15 protocols using prim 1640
1
Cellular Uptake of DNA Nanostructures
2
Esophageal Cancer Cell Lines Protocol
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ESCC cell lines, YES2, KYSE70, KYSE30, KYSE450, KYSE140, KYSE150, KYSE410, KYSE180, KYSE510, and COLO680N, were from professor Yutaka Shimada (Kyoto University, Kyoto, Japan) as gifts. All these cell lines were cultured in PRIM1640 (Invitrogen) with 10% FBS, except KYSE150, which was cultured in a 1:1 mixture of RPMI1640 and F12 (Gibco, Thermo Fisher Scientific) culture medium with 2% FBS. Immortalized esophageal epithelium cell line, NE3, was donated by professor Enmin Li (Shantou University, Shantou, Guangdong, P.R. China) and was cultured in a 1:1 mixture of EpiLife and dKSFM (Gibco, Thermo Fisher Scientific). All cell lines were used within five passages from thawing for experiments, and were tested for Mycoplasma contamination using Mycoplasma Real-time PCR Detection Kit (Applied Biosystems). All cell lines were authenticated using short tandem repeat profiling by Beijing Microread Genetics Co. Ltd. All of these cells were maintained at 37 C with 5% CO 2 .
3
Culturing MDA-MB-231 Breast Cancer Cells
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Human breast cancer cell MDA-MB-231 was purchased from Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, and cultured with PRIM 1640 (Gibco Life Technologies, Grand Island, NY, USA), 10% fetal bovine serum (FBS, ExCell Bio Inc., Australia) and 1% Penicillin Streptomycin at 37°C with 5% CO2 in a humidified cell incubator under 95%/5% (v/v) mixture of air and CO2.
4
Caco-2 and HCT116 cell culture protocol
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Caco-2 human intestinal epithelial cells and HCT116 human colorectal cancer cells were cultured in Gibco PRIM 1640 and DMEM media, respectively, supplemented with 10% fetal bovine serum (FBS), penicillin, and streptomycin. Cells were maintained in a 5% CO2 incubator at 37°C and were routinely tested to exclude mycoplasma contamination. During the study, Caco2 cells were stimulated with RTS (600 µmol/L) and LPS (10 µg/mL) and HCT116 cells were stimulated with RTS (600 µmol/L) and LPS (5 µg/mL) for 24h. Controls were treated with the same amount of vehicle.
5
Cell Line Culture for NSCLC Research
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The NSCLC cell lines A549, A549/DDP were purchased from Cell Bank of Chinese Academy of Sciences (Shanghai, China). A549 and A549/DDP Cells were cultured in PRIM 1640 (GIBCO, NY, USA) containing 10% fetal bovine serum (Biological Industries) and 1% penicillin/streptomycin at 37℃ under 5% CO2.
6
Lung Cancer Cell Line Cultivation
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The lung cancer cell lines A549, H460, H358, and H1299 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). In addition, A549-Luc-Puro-shRNA-hYAP1-Neo, A549-Luc-Puro-shRNA-Neocells were purchased from Biowit Technologies (Shenzhen, China). Cell were cultured in F12K (GIBCO, NY, USA), PRIM 1640 (GIBCO, NY, USA), or DMEM (GIBCO, NY, USA) supplemented with 10% fetal bovine serum (FBS, BI) and 1% penicillin/streptomycin at 37 °C and 5% CO2. The expression plasmids encoding YAP1, YAPS94A, TEAD, Slug-Luc, and mSlug-Luc were constructed using PGL3; verteporfin was purchased from Sigma.
7
Maintaining Human Gastric Cell Lines
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Human gastric mucosal epithelial cell line GES‐1, gastric cancer cell lines SNU‐216, AGS, SGC‐7901 and BGC‐823 cells were obtained from Shanghai Institute of Cell Biology (Shanghai, China) and maintained in PRIM‐1640 (GIBCO, Rockville, MD, USA) with 10% fetal bovine serum (GIBCO), 100 U/mL penicillin, and 100 μg/mL streptomycin in humidified incubator with 5% CO2 at 37°C.
8
Culturing Cell Lines for Ferroptosis Study
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Human BC T24, 5637, J82, and UM-UC3 cell lines were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells (except for 5637 cells) were cultured with Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Waltham, MA, USA; C11995500BT; Thermo Fisher, Waltham, MA, USA) supplemented with 10–20% fetal bovine serum (FBS; Gibco, Waltham, MA, USA; 10099-141c; Thermo Fisher, Waltham, MA, USA). The 5637 cells were cultured with PRIM1640 (Gibco, Waltham, MA, USA; C11875500BT; Thermo Fisher, Waltham, MA, USA). All cells were housed in a humidified atmosphere at 37 °C with 5% CO2. The ferroptosis inhibitor ferrostatin-1 and ferroptosis activator RSL3 were purchased from Med ChemExpress (MCE).
9
Culturing Human Liver Cell Lines
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All cell lines were purchased from the company (Shanghai Zhong Qiao Xin Zhou Biotechnology Co., Ltd., Shanghai, China) and were cultured under an atmosphere of 5% CO2 at 37 °C as described previously [19 (link)]. The human HCC cell lines (HCCLM3, HepG2 and Huh7) and LO2 were cultured in Dulbecco’s modified Eagle medium (DMEM; Gibco, Carlsbad, CA, USA) medium, PRIM‐1640 (Gibco), respectively. The medium was supplemented with 10% FBS (Gibco), penicillin (100 U·mL−1) and streptomycin (100 mg·mL−1).
10
In Vitro Cell Culture and Transfection
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The NCI-H1650 and NCI-H1299 lines were purchased from the Science Cell Laboratory. Cells were cultured in PRIM 1640 (GIBCO, USA) supplemented with 10 % fetal bovine serum (Cromwell, USA) and 100 μL/mL penicillin and streptomycin (Sigma-Aldrich, USA) and placed at 37° C with 5% CO2. 2 μg plasmid or 500 nM si-RNA/microRNA/antisense morpholino oligonucleotide (AMO)-microRNA or its NC was transfected into cells with LipofectamineTM 2000 (Invitrogen, Carlsbad, CA, USA), respectively. 10 μM cisplatin was added into medium for cisplatin treatment. As for co-culture assay, we used transwell membranes in 12-well plates. CAFs or NFs were plated into the upper chamber, and NCI-H1650 and NCI-H1299 was cultured in lower chamber.
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