Surgical masks were purchased from Nanchang Zhenfeng Sanitary Materials Co., Ltd. Copper (II) chloride dihydrate (CuCl2·2H2O), trisodium citrate, polyvinyl pyrrolidone (PVP, K30), sulfuric acid (H2SO4, 98%), and sodium sulfide (Na2S·9H2O) were brought from Sinopharm Chemical Reagent Co., Ltd. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and phosphate-buffered saline (PBS) were obtained from Beyotime Biotechnology. The adhesion promoter (CS-555) containing the amino-contained silane coupling agent was purchased from Nanjing Chuangshi Chemical Additives Co., Ltd. All the chemicals were used as received without further purifications. Infrared lamp (Philips, T04-R95E100W) was purchased from Shanghai Yafu Lighting Appliance Co., Ltd.
Phosphate buffered saline pbs
Manufactured by Beyotime
Sourced in China, United States
Phosphate-buffered saline (PBS) is a widely used buffer solution composed of sodium phosphate and sodium chloride. It is designed to maintain a stable pH and osmolarity, providing a physiologically compatible environment for biological samples. The primary function of PBS is to maintain the chemical and physical properties of cells, tissues, or other biological materials during various experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (3 mentions)
Ripa lysis buffer, by Beyotime (3 mentions)
Acetonitrile acn, by Tedia (2 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
Penicillin streptomycin, by Beyotime (2 mentions)
Annexin 5 fitc apoptosis detection kit, by Beyotime (2 mentions)
Bca protein assay kit, by Beyotime (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Beyoecl plus kit, by Beyotime (2 mentions)
Polyvinylidine fluoride membrane, by Beyotime (2 mentions)
Tris buffered saline with tween 20, by Beyotime (2 mentions)
Bovine serum albumin (bsa), by Beyotime (2 mentions)
Sodium dodecyl sulfate polyacrylamide gel electrophoresis, by Beyotime (2 mentions)
Trypsin, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium dmem, by Merck Group (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Dimethyl sulfoxide (dmso), by Beyotime (2 mentions)
Streptomycin, by Beyotime (2 mentions)
48 protocols using phosphate buffered saline pbs
1
Copper-Based Antimicrobial Mask Fabrication
2
Cell Viability Assay with siRNA Transfection
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A total of 1.5x10 4 cells were plated in 96-well plates in triplicate and were grown to 30-50% confluency at the time of transfection. The siRNA-Lipo-fectamine™ 2000 complex (50 µl) was administered to each well containing 100 µl DMEM without FCS. The medium was replaced with DMEM containing 10% (vol/vol) FCS at 6 h of transfection. MTT (20 µl; 5 mg/ml; Solarbio Technology) diluted in phosphate-buffered saline (PBS; Beyotime Institute of Biotechnology, Shanghai, China) was added to the medium at 24, 48 and 72 h after the transfection. After an incubation for 4 h, the medium was removed and the cells remained at the bottom of the wells. Dimethyl sulfoxide (200 µl; Solarbio Technology) was added to each well to dissolve the formazan crystals in the cells. The absorbance was measured using a microplate reader (MK3; Thermo Labsystems Inc., Beverly, MA, USA) at 540 nm to determine the quantities of viable cells. All experiments were performed in triplicate. Data were normalized to their respective controls and presented as a bar graph.
3
Quantitative Proteomic Analysis of FRα and FRβ
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Stable isotope-labeled amino acids were supplied by Cambridge Isotope Laboratories, Inc. (Andover, MA, USA). The synthetic proteolytic peptides and their corresponding stable isotope-labeled internal standards were synthesized by ChinaPeptides Co., Ltd. (Shanghai, China). FRα and FRβ were purchased from Abnova (Taipei, Taiwan) and Abcam (Cambridge, UK), respectively. Ammonium bicarbonate (NH4HCO3) was obtained from Qiangshun Chemical Reagent Co., Ltd. (Shanghai, China). DL-dithiothreitol (DTT), iodoacetamide (IAA), Tris-HCl and Triton X-114 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ethylenediaminetetraacetic acid disodium salt (EDTA-2Na) was obtained from Sinopharm Chemical Reagent Company (Shanghai, China). Sequencing grade modified trypsin was purchased from Promega (Madison, WI, USA) Phosphate buffered saline (PBS) was purchased from the Beyotime Institute of Biotechnology (Jiangsu, China). Acetonitrile (ACN) and methanol were obtained from Tedia Company, Inc. (Fairfield, OH, USA). Trifluoroacetic acid (TFA) and formic acid (FA) were purchased from Aladdin Chemistry Co., Ltd. (Shanghai, China) and Xilong Chemical Industrial Factory Co., Ltd. (Shantou, China), respectively. Sodium dodecyl sulfate (SDS) was obtained from Generay Biotech Co., Ltd (Shanghai, China). Water was purified and deionized with a Milli-Q system from Millipore (Bedford, MA, USA).
4
Exosome Isolation from Cell Culture Media
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ExoQuick precipitation kit (System Biosciences, Mountain view, CA, USA) was utilized for isolating exosomes from cell culture medium following the manufacturer’s instructions. In short, cell culture medium was collected and centrifuged (3000 × g, 15 min), followed by addition of ExoQuick precipitation solution to supernatant. After centrifugation (1500 × g, 30 min), exosome pellet was re-suspended in phosphate-buffered saline (PBS; Beyotime, Shanghai, China). Exosomes were visualized by transmission electron microscopy (TEM; JEOL, Akishima, Japan) and exosome protein markers (CD63 and CD9) were identified by WB assay.
5
Isolation and Characterization of Bacterial Lipopolysaccharides
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Bacterial lipopolysaccharide (LPS), concanavalin A (Con A), corn oil, and dexamethasone were purchased from Sigma-Aldrich (St Louis, USA). CCl
4 was purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Potassium luciferin (Gold Biotechnology, St Louis, USA) was dissolved in phosphate-buffered saline (PBS; Beyotime, Shanghai, China) at 15 mg/mL and stored at –20°C.
4 was purchased from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China). Potassium luciferin (Gold Biotechnology, St Louis, USA) was dissolved in phosphate-buffered saline (PBS; Beyotime, Shanghai, China) at 15 mg/mL and stored at –20°C.
6
Culturing ESCC Cell Lines with F. nucleatum
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Human ESCC cell lines KYSE-150 and ECA-109 obtained from Shanghai Cell Bank, Chinese Academy of Sciences were cultured in RPMI-1640 medium (Gibco, Gaithersburg, MD, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Gaithersburg, MD, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China) at 37°C in a humidified 5% CO2 atmosphere. The F. nucleatum strain ATCC 25,586 was purchased from the China General Microbiological Culture Collection Center (Beijing, China) and cultured in fastidious anaerobe broth (Tuopu biotechnology, Qingdao, China) at 37°C under anaerobic condition. For in vitro co-culture experiments, cells were washed with phosphate-buffered saline (PBS, Beyotime, Shanghai, China) and then incubated with F. nucleatum at a multiplicity of infection (MOI) of 100 in antibiotic-free RPMI-1640 medium supplemented with 10% FBS at 37°C under 5% CO2 for 24 h.
7
Ochratoxin A Cytotoxicity in Caco-2 Cells
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OTA powder (C20H18ClNO6; molecular weight, 403) was purchased from Pribolab (Qingdao, China). Human colon adenocarcinoma Caco-2 cells (passage number 18) were acquired from the American Type Culture Collection (Manassas, VA, USA). Fetal bovine serum (FBS) and Dulbecco’s modified Eagle’s medium (DMEM) were obtained from Gibco (Carlsbad, CA, USA). Nonessential amino acids (NEAA), trypsin (2.5%), antibiotics (100 units/mL penicillin, 100 µg/mL streptomycin), phosphate-buffered saline (PBS), Western and IP Cell Lysis Buffer, and an Annexin V-FITC apoptosis detection kit were supplied by Beyotime Biotechnology (Shanghai, China). A stock solution of OTA (1000 μg/mL) was obtained by dissolving OTA in methanol, and stored at −20 °C for later use. Rabbit anti-β-actin (58169S) and rabbit anti-MDM2 antibodies (86934S) were obtained from Cell Signaling Technology (Boston, MA, USA), and goat anti-rabbit IgG conjugated to horseradish peroxidase was obtained from Bioss (Beijing, China).
8
Oxidative Stress and Cell Viability Assays
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Glucose-high Dulbecco’s modified Eagle’s medium/F12 (DMEM/F12), glucose-free DMEM and fetal bovine serum (FBS) were purchased from Gibco (Grand Island, NY, USA); poly-L-lysine and H2O2 were from Sigma-Aldrich (Milan, Italy); 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazol-carbocyanine iodide (JC-1), Fluo-3, AM and Annexin V-FITC/propidium iodide (PI) were obtained from Molecular Probes, Invitrogen (Milan, Italy), and MTS was from Promega Corp. (Madison, WI, USA); Hank’s solution and trypsin were obtained from HyClone (Logan, UT, USA), and Hoechst 33342, phosphate-buffered saline (PBS) and penicillin/streptomycin (Pen/Strep) were from Beyotime (Shanghai, China). The lactate dehydrogenase (LDH) assay kit was purchased from Roche Molecular Biochemicals (Indianapolis, IN, USA).
9
Collagen I Immunohistochemistry in Myocardial Tissue
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Myocardial tissue samples were fixed with 4% polyformaldehyde, embedded in paraffin, cut into slices (thickness, 10 µm), dewaxed and hydrated. The slices were incubated with 3% hydrogen peroxide (Beyotime Institute of Biotechnology), washed with phosphate-buffered saline (PBS; Beyotime Institute of Biotechnology), blocked with 10% normal goat serum (Beyotime Institute of Biotechnology) at 37°C for 10 min, incubated at 37°C with rabbit polyclonal anti-collagen I (dilution 1:100) for 90 min, washed with PBS, incubated at 37°C with secondary antibodies (dilution 1:2,000) for 15 min, washed with PBS, incubated at 37°C with horseradish peroxidase-labeled streptavidin (Beyotime Institute of Biotechnology) for 20 min, washed with PBS, colored with color developing reagent (Beyotime Institute of Biotechnology), stained with hematoxylin (Beyotime Institute of Biotechnology) at 37°C for 20 min, dehydrated with alcohol and sealed with neutral resins.
10
CXCR4 Expression Analysis in Corneal Tissue
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Corneas were cut and fixed in 10% neutral buffered formalin (Beijing Yili Fine Chemicals Co., Ltd.; Beijing, China) for 24 h. Paraffin-embedded tissue sections (4 μm) were deparaffinized, rehydrated, and treated with 0.3% hydrogen peroxide in methanol (Beyotime Institute of Biotechnology, Inc., Haimen, China) for 30 min, to eliminate endogenous peroxidase activity. The tissue sections were incubated for 60 min at room temperature with rabbit anti-mouse CXCR4 polyclonal antibody (1:200 dilution; cat. no. ab2074; Abcam). Following three washes of 3 min with phosphate-buffered saline (PBS; Beyotime Institute of Biotechnology, Inc.), a 3,3′-diaminobenzidine detection kit (PV9000; ZSGB-BIO, Beijing, China) was used for CXCR4 staining, according to the manufacturer's protocols. Images were captured with the Leica DM4000B biological microscope equipped with a Leica DFC 550 digital camera and Leica Application Suite version 4.2.0 software (Leica Microsystems GmbH, Wetzlar, Germany).
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