Mouse embryonic small intestinal samples from different stages were carefully dissected and harvested from the body. All samples used for histological sectioning and subsequent staining were taken from duodenal sections. For paraffin section, samples were fixed overnight at 4° C temperature in 4% paraformaldehyde in PBS buffer (Invitrogen) and incubated in 70% ethanol solution after three times’ wash with PBS buffer for paraffin section, and then were submitted to the Molecular Pathology and Imaging Core of the Center for Molecular Studies in Digestive and Liver Diseases (P30 DK050306) for further process, embed and sectioning for sectioning at tum thickness and then sectioned slices were kept at room temperature. For frozen section, samples were incubated overnight in 30% sucrose in PBS buffer after overnighted fixation at 4°C in 4% paraformaldehyde in PBS buffer (Invitrogen) until the intestinal tissues completely sink to the tube bottom, and then were embed in OCT for quick-frozen and stored at −80°C. OCT-embedded samples were performed the cryo-sectioning though a using a cryostat (Cryostar NX50, ThermoFisher Scientific) at 10 μm, dried for 10 minutes at room temperature before the further immunofluorescent staining or stored at −80°C.
Pbs buffer
Manufactured by Thermo Fisher Scientific
Sourced in United States, France, China, Austria, Australia, Switzerland, United Kingdom
PBS buffer is a commonly used buffer solution that maintains a physiologically relevant pH and ionic strength. It is composed of sodium phosphate and sodium chloride, and is primarily used to maintain the stability and activity of biological samples in laboratory applications.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (14 mentions)
Dmem high glucose media, by Thermo Fisher Scientific (8 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (8 mentions)
Pen strep, by Thermo Fisher Scientific (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (8 mentions)
Formic acid, by Thermo Fisher Scientific (7 mentions)
Binding buffer, by Thermo Fisher Scientific (6 mentions)
Propidium iodide, by Thermo Fisher Scientific (6 mentions)
Annexin 5, by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Bovine serum albumin (bsa), by Merck Group (6 mentions)
Trypsin, by Thermo Fisher Scientific (6 mentions)
Acetonitrile, by Thermo Fisher Scientific (6 mentions)
Streptomycin, by Thermo Fisher Scientific (5 mentions)
Agilent 6230 lc tof mass spectrometer, by Agilent Technologies (4 mentions)
Tribute peptide synthesizer, by Protein Technologies (4 mentions)
Axio observer a1 inverted microscope, by Zeiss (4 mentions)
Sp5 confocal fluorescence microscope, by Leica (4 mentions)
Facsaria cell sorter, by BD (4 mentions)
Waring blender, by Merck Group (4 mentions)
120 protocols using pbs buffer
1
Mouse Intestinal Tissue Preparation
2
Small Molecule and Peptide Synthesis Protocol
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Chemical reagents for small molecule and peptide synthesis were purchased from various vendors and used as received. The phospholipids were purchased from Avanti Polar Lipids (Alabaster, Al). PBS buffer, DMEM/High glucose media, RPMI 1640 media, and Pen/Strep were purchased from Thermal Scientific (Amarillo, TX). The Gram-positive bacteria (B. subtilis (ATCC 663) and S. aureus (ATCC 6538)) were purchased from Microbiologics (Cloud, MN) as lyophilized cell pellet. E. coli (BL 21) was a gift from the lab of Professor Mary F. Roberts at Boston College. NMR data of the small molecules were collected on a VNMRS 500 MHz NMR spectrometer. MS data were generated by using an Agilent 6230 LC TOF mass spectrometer. Peptide synthesis was carried out on a Tribute peptide synthesizer from Protein Technologies. The fluorescence anisotropy experiments were performed by using a SpectraMax M5 plate reader. Fluorescence images were taken on a Zeiss Axio Observer A1 inverted microscope. Confocal images were taken on the Leica SP5 confocal fluorescence microscope housed in the Biology Department of Boston College. Flow cytometry analyses were carried out on a BD FACSAria cell sorter also housed in the Biology Department of Boston College.
3
Small Molecule and Peptide Synthesis Protocol
4
Antimicrobial Efficacy Evaluation Protocol
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Norfloxacin and Levofloxacin were purchased from Energy Chemical. Moxifloxacin Hydrochloride was purchased from TCI. Ltd. PBS buffer, DMEM (Dulbecco's modified Eagle medium) and fetal bovine serum (FBS) were purchased from Termo Fisher Scientific (Shanghai, China). MCF-7 cells and HeLa cells were obtained from cell culture center of Institute of Basic Medical Sciences, Chinese Academy of Medical Science (Beijing, China). S. aureus ATCC 6538 and Candida albicans (C. albicans) ATCC 10231 were obtained from China General Microbiological Culture Collection Center. The Amp Escherichia coli (E. coli) TOP 10 was purchased from Beijing Bio-Med Technology Development Co.
5
Two-Photon Imaging of Yeast Cells
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Two-photon imaging microscopy was performed to obtain 2D representations of the two Saccharomyces cerevisiae strains. The images were collected on a Nikon A1-MP scanning microscope equipped with a Plan APO IR × 60 objective (NA, 1.27; Water Immersion, Nikon, Tokyo, Japan) at a scanning speed of one frame per second. An IR laser (Chameleon, Coherent, Palo Alto, CA, USA) was used to provide excitation at 820 nm. Fluorescence emission was collected on two detection channels: FF01-492/SP (400–492 nm) and FF03-525/50 (500–550 nm) (Semrock, New York, USA). The images provided in this article were obtained by merging these two detection channels.
Samples of the culture were collected and centrifuged at 8000× g for 5 min at 20 °C. The culture supernatant was removed, and the pellet was resuspended in PBS buffer (137 mM NaCl, 2.7 mM KCl, and 11.9 mM phosphate, pH 7.2) (ThermoFisher Scientific, Illkrich, France). Five microliters of cell suspension were placed between a glass slide and a coverslip for observation.
Samples of the culture were collected and centrifuged at 8000× g for 5 min at 20 °C. The culture supernatant was removed, and the pellet was resuspended in PBS buffer (137 mM NaCl, 2.7 mM KCl, and 11.9 mM phosphate, pH 7.2) (ThermoFisher Scientific, Illkrich, France). Five microliters of cell suspension were placed between a glass slide and a coverslip for observation.
6
Yeast Viability Determination by Flow Cytometry
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Yeast viability in preculture and during fermentation was determined via flow cytometry. The fluorochrome used was Propidium Iodide (PI) (Invitrogen, Molecular Probes, ThermoFisher Scientific, Illkrich, France) dissolved in filtered milliQ water at a concentration of 0.1 mg mL−1 [4 (link)]. Here, 1 µL of PI was added to 100 µL of diluted yeast suspension in PBS buffer (ThermoFisher Scientific, Illkrich, France). Samples were incubated in the dark 15 min before measurement.
7
Rat Model for Cardiovascular Research
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Male Sprague Dawley (SD) rats (weighting 250–320 g, purchased from Charles River, Wilmington, MA, USA) were used in this study. The rats were housed under controlled conditions with a 12-h light/dark cycle. Rat chow and water were provided ad libitum. All protocols were approved by the North Dakota State University Institutional Animal Care and Use Committee (IACUC) and executed in accordance with the Guide for the Care and Use of Laboratory Animals (National Institutes of Health, Bethesda, MD, USA). In this study, the osmotic minipumps were obtained from ALZET.com. Saline and PBS buffer were purchased from Thermo Fisher, Waltham, MA, USA. Ang II, Apelin-13, and other agents were purchased from Millipore Sigma, St. Louis, MO, USA.
8
Synthesis and Characterization of Photosensitive Nanoparticles
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Hematoporphyrin IX dihydrochloride (HPIX) was purchased from Frontier Scientific. Tetraethyl orthosilicate (TEOS), 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), Trypan Blue solution, silver nitrate, and absolute anhydrous ethanol were from Sigma Aldrich. Ammonium nitrate, nitric acid (68%), formaldehyde solution (37%), cetyltrimethylammonium bromide (CTAB), sodium hydroxide, sodium cyanide, Penicillin-Streptomycin (10,000 U/mL), DAPI (4′,6-diamidino-2-phenylindole), and PBS buffer were from Thermo Fisher. All chemicals were used as received. Human cervical cancer cells Hela (ATCC CCL-2) were obtained from American Type Culture Collection (ATCC). Dulbecco’s Modified Eagle’s Medium (DMEM) was purchased from Sigma, and fetal calf serum (FBS) from Hyclone.
9
Comprehensive Materials and Methods
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N-acetylcysteine (NAC) was obtained from Shanghai Aladdin Biochemical Technology Co., Ltd. (China). Soybean phosphatidylcholine (SPC), cholesterol (Chol), sodium deoxycholate (SDC) and lipopolysaccharides (LPS) were purchased from Shanghai Macklin Biochemical Co., Ltd. (China). 1, 2-Dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS) was available from Corden Pharma Switzerland LLC. Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum (FBS), L-glutamine, sodium pyruvate, PBS buffer and penicillin-streptomycin (P/S) were obtained from ThermoFisher Scientific Inc. (USA). All other solvents were of analytical grade or specific HPLC grade. Recombinant murine M-CSF was purchased from PeproTech, Inc. (USA). Bleomycin (BLM) was obtained from Hanhui Pharmaceuticals Co., Ltd. (China).
10
Rapid SARS-CoV-2 Antibody Detection Assay
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Tetraethyl orthosilicate (TEOS), 2-(N-morpholino) ethanesulfonic (MES), polyethyleneimine (PEI, 25 K), polyvinylpyrrolidone (PVP, 40 K), Tween 20, N-hydroxysuccinimide (NHS), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB), sodium borohydride (NaBH4), trisodium citrate (Na3C6H5O7, TSC), goat anti-human IgM, goat anti-human IgG, goat anti-rabbit IgG, bovine serum albumin (BSA) were obtained from Sigma-Aldrich (USA). SARS-CoV-2 S protein (recombinant) and S protein antibody (Ab) were obtained from Sino Biological Inc. (Beijing, China). Chloroauric acid tetrahydrate, sodium borohydride, formaldehyde (37 %, w/w), ammonia solution (28 %, w/w), Chloroauric acid tetrahydrate (HAuCl4·4H2O), and silver nitrate (AgNO3) were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). PBS buffer (10 mM, pH 7.4) and fetal bovine serum (FBS) were obtained from Thermo Fisher (Shanghai, China). Nitrocellulose (NC) membrane was purchased from Sartorius (UniSart CN95 and CN140, Spain) and Millipore Corporation (HF135, USA), respectively. The sample pad, absorbent pad, conjugate pad, and PVC bottom plate were obtained from Jieyi Biotechnology Co (Shanghai, China). All chemicals were of analytical grade and were utilized as received unless mentioned otherwise.
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