4-Hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (Tempol, purity ≥ 98%) was obtained from Aladdin Biochemical Technology Co., Ltd (Shanghai, China). l -α-Glycerophosphocholine (GPC, purity ≥ 99%), succinic anhydride (SA, purity ≥ 98%), cholesterol (Chol, purity ≥ 98%), N,N′-carbonyldiimidazole (CDI, purity ≥ 99%), 4-dimethylaminopyridine (DMAP, purity ≥ 98%) and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, purity ≥ 99%) were purchased form DB Technology Co., Ltd (Shanghai, China). RPMI medium 1640, fetal bovine serum (FBS), methyl tetrazolium (MTT), 4′,6-diamidino-2-phenylindole (DAPI) and penicillin–streptomycin solution were supplied by Beyotime (Shanghai, China). Lipopolysaccharide (LPS), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Oil Red O (ORO) were purchased from Sigma-Aldrich (MO, USA). The chemicals and solvents were provided from the domestic suppliers and used without further purification, except where specified below.
Rpmi 1640 medium
Manufactured by Beyotime
Sourced in China, United States
RPMI-1640 medium is a commonly used cell culture medium formulated to support the growth and maintenance of a variety of cell types, including human and animal cell lines. It provides a balanced mixture of nutrients, vitamins, and other essential components required for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Beyotime (8 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Cell counting kit 8 cck 8, by Beyotime (2 mentions)
Biotek synergy 2, by Agilent Technologies (2 mentions)
Penicillin streptomycin, by Beyotime (2 mentions)
Penicillin, by Beyotime (2 mentions)
Dulbecco modified eagle medium (dmem), by Beyotime (2 mentions)
Oil red o, by Merck Group (1 mentions)
Penicillin streptomycin solution, by Beyotime (1 mentions)
Diclofenac sodium, by Maclin Biochemical Technology (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Dulbecco s modified eagle medium dmem, by Solarbio (1 mentions)
Vantage se, by BD (1 mentions)
Cd44 fluorescein isothiocyanate fitc, by BD (1 mentions)
Transwell apparatus, by Corning (1 mentions)
Phosphate buffer saline (pbs), by Beyotime (1 mentions)
Calcein am staining, by Beyotime (1 mentions)
Cell counting kit 8 solution, by Beyotime (1 mentions)
Amiloride, by Merck Group (1 mentions)
25 protocols using rpmi 1640 medium
1
Synthesis and Characterization of Tempol-Loaded Liposomes
2
Anti-inflammatory Compound Evaluation Protocol
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Dexamethasone sodium phosphate (DSP, purity ≥ 99%) and diclofenac sodium (DS, purity ≥ 99%) were purchased from Macklin Biochemical Technology Co., Ltd. (Shanghai, China). Calcium chloride (CaCl2, purity ≥ 98%) and Ethylene diamine tetraacetic acid (EDTA, purity ≥ 98%) were purchased from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Lipopolysaccharide (LPS) was purchased from Sigma Co., Ltd. (St. Louis, MO, USA). Complete Freund’s Adjuvant (CFA) with 10 mg/mL of heat-killed mycobacterium tuberculosis was supplied by Chondrex Co., Ltd. (Redmond, WA, USA). Biochemical reagents including Cell Counting kit-8 (CCK8), RPMI medium 1640, fetal bovine serum, and 4′,6-diamidino-2-phenylindole (DAPI) were purchased from Beyotime (Shanghai, China). Nitric oxide (NO) kit, ELISA kit for analysis of tumor necrosis factor-a (TNF-a), and interleukin-6 (IL-6) were supplied by KeyGEN BioTECH (Nanjing, China). Alanine transaminase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (Crea) kits were supplied by Nanjing Jiancheng Co., Ltd. (Nanjing, China). Nuclear factor kappa-B p65 (NF-κB p65), phospho-NF-κB p65 (p-NF-κB p65), cyclooxygenase-2 (COX-2), and inductible nitric oxide synthase (iNOS) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).
3
Zebrafish Tumor Cell Inoculation
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Zebrafish tumor models were developed as previously described [22 (link)]. Animals were housed in the Laboratory Zebrafish Center at Nantong University. For tumor cell inoculation, 300 tumor cells in 5 nL RPMI 1640 medium labeled with 2 g/mL DiI (Beyotime, C1036) were injected into the perivitelline cavity of zebrafish embryos at 48 h post-fertilization using a microinjection system (WPI, Sarasota, Florida, USA).
4
CCK-8 Cytotoxicity Assay in Gastric Cancer Cells
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Stably transfected AGS and HGC-27 cells (2×103 cells/well) were seeded in 96-well plates and cultivated for 24, 48, 72 or 96 h. Then, 10 µl cholecystokinin octapeptide (CCK-8) reagent [10% (v/v) in serum-free RPMI-1640 medium; Beyotime Biotechnology] was added to each well and incubated at 37°C for 1 h. The absorbance at 450 nm was measured using a microplate reader (BioTek Synergy 2; BioTek Instruments Inc., Winooski, VT, USA).
5
Culturing Hepatocellular Carcinoma Cell Lines
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The hepatocellular carcinoma cell lines Hep G2, Hep 3B, and Huh-7 were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and the SMMC-7721 cell line was a kind gift from professor Ye (Zhengzhou University, China). The cells were incubated in RPMI-1640 medium (Beyotime Institute of Biotechnology, Shanghai, China) or Dulbecco's Modified Eagle Medium (DMEM, Solarbio Life Science, Beijing, China) supplemented with 10% fetal bovine serum (FBS; HyClone, Utah, USA), 100 U/mL penicillin, 100 mg/L streptomycin in an incubator (Thermo Fisher Scientific, USA) at 37° C and 5% CO2. The cells used for experiments were in the exponential growth phase.
6
CD44 and CD24 Expression Analysis
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Cells (1×106/100 ml) were blocked with RPMI-1640 medium containing 3% BSA (Beyotime Institute of Biotechnology) for 30 min on ice. Cells were subsequently incubated with the following primary antibodies: CD44-fluorescein isothiocyanate (FITC) and CD24-phycoerythrin (PE; dilution, 1:100 dilution for 106 cells/100 ml; BD Biosciences, San Jose, CA, USA). Cells were subsequently washed using PBS with 3% BSA. Analysis was performed using a fluorescence-activated cell sorting (FACS) Vantage SE flow cytometer (CellQuest Pro version 6.1 software; BD Biosciences).
7
Cell Migration and Invasion Assays
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A total of 1 × 105 cells were seeded onto a fibronectin-coated polycarbonate membrane insert in a transwell apparatus manufactured by Corning (NY, USA), with a pore size of 0.8 mm. In the lower chamber of the transwell, 600 μl of RPMI 1640 medium supplemented with fetal bovine serum from Beyotime Institute of Biotechnology was added as a chemoattractant. Following a 12-hour incubation, the insert was carefully washed with PBS to remove any non-adherent cells from the upper surface. Next, the GC cells that migrated through the membrane and attached to the lower surface of the insert were fixed using 4% formaldehyde. To visualize and quantify the migrated cells, the fixed cells were stained with a 0.2% crystal violet solution obtained from Shanghai Qiaoxing Trading Corporation in Shanghai, China. Cell counts were determined using ImageJ software, and photographs were captured. The Matrigel invasion assay was conducted following a procedure similar to the cell migration assay described above. However, in the Matrigel invasion assay, the transwell membrane was pre-coated with ECMatrix™, and the cells were incubated for 14 hours. Each experiment was repeated a minimum of three times.
8
Culturing Lung Cell Lines
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A human normal lung cell line (2B) and two LUAD cell lines (A549 and H1650) were purchased from Beyotime Biotechnology (Procell, Wuhan, China) and cultured in RPMI-1640 medium containing 10% foetal bovine serum at 37 °C in 5% CO2. All media and supplements were purchased from Invitrogen (Carlsbad, CA, USA).
9
Lipoic Acid, Phytic Acid, and Sodium Bicarbonate Protocol
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Lipoic acid (LA), phytic acid (PA) and sodium bicarbonate (NaHCO3) were purchased from Macklin. RPMI 1640 medium, fetal bovine serum (FBS), penicillin–streptomycin (PS), Calcein AM staining, Cell counting Kit-8 (CCK-8) and phosphate buffer saline (PBS) were provided by Beyotime Biotechnology (Shanghai, China).
10
Cytotoxicity Assay for DOX and SML
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The cells were seeded onto 96-well plates at a density of 2,000 cells/well. Following treatment with 1, 0.1 µg/ml DOX for 1, 2, 3 and 4 days; 2, 25, 50, 75 and 100 µg/ml SML for 1, 2, 3 and 4 days; 3, DOX alone or SML plus DOX for 1, 2, 3 and 4 days, the cells were washed with PBS. RPMI-1640 medium (100 µl) and 10 µl cell counting kit-8 solution (Beyotime Institute of Biotechnology) were added to each well for another 2 h at 37°C. The optical density value was read at 450 nm using an ultraviolet spectrophotometer.
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