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Uranyl formate

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Uranyl formate is a chemical compound that is commonly used as a negative stain in transmission electron microscopy (TEM) for the visualization and analysis of biological samples. It serves as a contrast-enhancing agent, providing improved contrast and visibility of the sample under the electron beam.

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Lab products found in correlation

Tecnai t12, by Thermo Fisher Scientific (6 mentions) Pcr tubes, by Avantor (3 mentions) Carbon formvar grids, by Electron Microscopy Sciences (3 mentions) Amicon ultra filtration devices, by Thermo Fisher Scientific (3 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (3 mentions) Tween 20, by Merck Group (3 mentions) Tris base, by Merck Group (3 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (3 mentions) Glycerol, by Merck Group (3 mentions) Agarose, by Lonza (3 mentions) Glutaraldehyde, by Merck Group (2 mentions) Triton x 114, by Merck Group (2 mentions) Magnesium chloride, by Merck Group (2 mentions) Endotoxin test cartridges, by Charles River Laboratories (2 mentions) Accugene 10 tbe buffer, by Avantor (2 mentions) Seton ultracentrifugation tubes, by Thermo Fisher Scientific (2 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions) Sodium chloride (nacl), by Merck Group (2 mentions)

36 protocols using uranyl formate

1

Synthesis and Characterization of Selenium Nanoparticles

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Reagents for the synthesis of SeNPs: chitosan, bovine serum albumin, sodium selenite, ascorbic, and acetic acid were purchased from Sigma-Aldrich. Reagents for electron microscopy: glutaraldehyde, cacodylate and formaldehyde were purchased from Sigma. Uranyl formate was purchased from Electron Microscopy Sciences (Hatfield, PA, United States).
Estevez H., Palacios A., Gil D., Anguita J., Vallet-Regi M., González B., Prados-Rosales R, & Luque-Garcia J.L. (2020). Antimycobacterial Effect of Selenium Nanoparticles on Mycobacterium tuberculosis. Frontiers in Microbiology, 11, 800.
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2

Purification of TrpLE-MSP1E3D1 Fusion Protein

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The gene encoding the TrpLE protein fused to MSP1E3D1 was prepared by Gene Universal (Newark, DE). Uranyl formate and 400-mesh copper grids were purchased from Electron Microscopy Services (Hatfield, PA). All other reagents were of standard laboratory grade.
Julien J.A., Mutchek S.G., Fernandez M.G, & Glover K.J. (2021). Facile production of tagless membrane scaffold protein for nanodiscs. Analytical biochemistry, 638, 114497.
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3

Liposome Preparation and Characterization

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Lipids DOPC and DOPS, and PEG-PE, cholesterol and rhodamine-PE, the mini-extruder, extrusion membranes, and accessories were purchased from Avanti Polar Lipids. Endotoxin test cartridges (0.05–5.0 EU/mL) were purchased from Charles River. Accugene 10× TBE buffer, PCR tubes, and 96-well PCR plates (Axygen) were purchased from VWR. SYBR Safe and PicoGreen stains were purchased from Life Technologies Corporation. Agarose and cell lysis buffer were purchased from Lonza. Glycerol, Tris base, EDTA, Triton X-114, octyl β-d-glucopyranoside (OG), Tween20, magnesium chloride, magnesium sulfate, sodium chloride, and paraformaldehyde were purchased from Sigma-Aldrich. RPMI, PBS, FBS, and penicillin-streptomycin were purchased from Gibco. Carbon Formvar grids and uranyl formate were purchased from Electron Microscopy Sciences. The 96- and 384-well fluorescence assay plates were purchased from BD Biosciences. Amicon Ultra filtration devices, Optiprep media, and Seton ultracentrifugation tubes were purchased from Fisher Scientific.
Perrault S.D, & Shih W.M. (2014). Virus-Inspired Membrane Encapsulation of DNA Nanostructures To Achieve In Vivo Stability. ACS Nano, 8(5), 5132-5140.
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4

Peptide Synthesis and Conjugation Reagents

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Rink-amide resin, protected amino acids, and solvents used for peptide synthesis were obtained from Protein Technologies Inc. (Tucson, AZ). Fluorescein isothiocyanate (FITC), trifluoroacetic acid, thioanisole, anisole, 1,2-ethanedithiole, methyl-tert-butyl ether, N,N-diisopropylethylamine, hydrazine, GSH, MA, and 3MA were purchased from Sigma-Aldrich (Saint-Louis, MO). Sulfosuccinimidyl 4-[N-maleimidomethyl]cyclohexane-1-carboxylate (sulfo-SMCC) was obtained from Novachem (Calgary, Canada) and p-SNC-deferoxamine (DFO) was from Macrocyclics Inc. (Plato, TX). AlDoxorubicin was purchased from Medkoo Biosciences (Chapel Hill, NC). Doxorubicin was from LC Laboratories (Woburn, MA), and N2′- deacetyl-N2′-(3-mercapto-1-oxopropyl)-maytansine was supplied by Carbosynth Ltd (Compton, WB). Uranyl formate was purchased from Electron Microscopy Sciences (Hatfield, PA), and 4′6-diaminidino-2-pheylindole (DAPI) and LysoTracker Red were purchased from Life Technologies Inc. (Norwalk, CT). Cyanine5.5 NHS ester was obtained from Lumiprobe Corporation (Hallandale Beach, FL) and luciferin was supplied by Caliper LifeSciences (Hopkinton, MA).
Bellat V., Ting R., Southard T.L., Vahdat L., Molina H., Fernandez J., Aras O., Stokol T, & Law B. (2018). Functional Peptide Nanofibers with Unique Tumor Targeting and Enzyme-Induced Local Retention Properties. Advanced functional materials, 28(44), 1803969.
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5

Negative Staining of Glycoprotein

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Low and high fractions were sorted directly on glow-discharged carbon-coated TEM grids (Electron Microscopy Sciences). Samples were then incubated with the uncoupled mouse monoclonal antibody raised against GPC (10 μg ml−1; GD01-AG02; NR-2565; BEI resources), followed by a secondary Rabbit anti-mouse antibody (MP Biomedicals) and revealed by Protein A conjugated to 10 nm gold beads. Negative staining was achieved by incubating samples in Uranyl formate (Electron Microscopy Sciences) for 30 sec. The grids were examined on a JEOL 1200EX Transmission electron microscope and images were recorded with an AMT 2k CCD.
Gaudin R, & Barteneva N.S. (2015). Sorting of small infectious virus particles by flow virometry reveals distinct infectivity profiles. Nature communications, 6, 6022.
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6

Oligolysine-PEG Nanoparticle Preparation

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Tris/Borate/EDTA buffer, PCR tubes and 96-well PCR plates were purchased from VWR. Oligolysine (K4–K10 and R6) were purchased from Peptide 2.0 as crude in a 5 mg scale. Oligolysine K10–PEG1K, K10–PEG5K and K10–PEG20K were purchased from Alamada polymers in 100 mg scale. Agarose was purchased from Lonza. Magnesium chloride, sodium chloride, glycerol, Tris base, EDTA and Tween20 was purchased from Sigma-Aldrich. RPMI, PBS, FBS and penicillin–streptomycin were purchased from Gibco. Carbon Formvar grids and uranyl formate were purchased from Electron Microscopy Sciences. Amicon Ultra filtration devices and 3.5 K MWCO Slide-A-Lyzer mini dialysis devices were purchased from Fisher Scientific. DNA gel extraction spin column was purchased from Bio-Rad.
Ponnuswamy N., Bastings M.M., Nathwani B., Ryu J.H., Chou L.Y., Vinther M., Li W.A., Anastassacos F.M., Mooney D.J, & Shih W.M. (2017). Oligolysine-based coating protects DNA nanostructures from low-salt denaturation and nuclease degradation. Nature Communications, 8, 15654.
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7

Ultrastructural Analysis of Tau Aggregates

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Electron microscopy of negatively stained Tau aggregates is performed using 600-mesh carbon-coated copper grids (SPI). The samples were incubated for 30 seconds on a glow-discharged grid, and then the solution was removed by filter paper. Three washing steps with double distilled H2O were followed by three staining steps with 0.75% (w/v) uranyl formate (Electron Microscopy Sciences). The samples were imaged using a Talos L120C operated at 100 keV. Micrograph images were recorded using a 4kx 4k Thermofisher Scientific Ceta CMOS Camera.
Frostegård Å., Courtois S., Ramisse V., Clerc S., Bernillon D., Le Gall F., Jeannin P., Nesme X, & Simonet P. (1999). Quantification of Bias Related to the Extraction of DNA Directly from Soils. Applied and Environmental Microbiology, 65(12), 5409-5420.
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8

Native Gel Electrophoresis and Electron Microscopy of hTRiC

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Clear native gel electrophoresis was performed using a 4–16% Bis–Tris (pH 7.4) gel system from Invitrogen per manufacturer instructions. The gel was stained with GelCode blue stain reagent (Thermo Fisher Scientific). The proteinase K (PK) conformational cycling and radiolabeled actin folding assays were performed as described previously10 (link). For negative stain electron microscopy, hTRiC was diluted 10 times to 0.05 mg ml−1 in 20 mM HEPES, 50 mM NaCl, 5 mM MgCl2, 1 mM EDTA, 1 mM DTT. 5 μl sample was placed for 1 min on a formvar/carbon coated 200 holey mesh Ultra Thin copper grid (FCF200-Cu-UA, Electron Microscopy Sciences). The grid was blotted on filter paper, floated over stain for 1 min in 1% uranyl formate made fresh from powder (Electron Microscopy Sciences), and air dried. Grids were imaged using a JEOL JEM1400 transmission electron microscope.
Collier M.P., Moreira K.B., Li K.H., Chen Y.C., Itzhak D., Samant R., Leitner A., Burlingame A, & Frydman J. (2021). Native mass spectrometry analyses of chaperonin complex TRiC/CCT reveal subunit N-terminal processing and re-association patterns. Scientific Reports, 11, 13084.
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9

Structural Analysis of Human Dicer Enzyme

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A 3-μL drop of 50 nM human WT Dicer in 20 mM HEPES, pH 7.5, 150 mM KCl, 3 mM EDTA, 1 mM DTT, and 2.5% glycerol was applied onto a carbon-coated copper grid previously glow-discharged using an ELMO glow discharge system (Cordouan Technologies, France). After 1 min, excess liquid was blotted and stained for 1 min with 1.5% freshly prepared uranyl formate (Electron Microscopy Sciences, PA). Samples were imaged using a FEI Tecnai T12 (Eindhoven, The Netherlands) Transmission Electron Microscope (TEM) equipped with a LaB6 filament and operated at an acceleration voltage of 120 kV. Micrographs were collected at defocus between 1 and 3 μm on a FEI Eagle 4 k × 4 k CCD camera at a magnification of ~ 67,000×. From these micrographs, 125,000 particles were extracted and 2D align using xmipp from Scipion [54 (link)]. The particles were classified in a minimum of 2 classes and the 2D volumes were measured using ImageJ [55 (link)].
Bouvette J., Korkut D.N., Fouillen A., Amellah S., Nanci A., Durocher Y., Omichinski J.G, & Legault P. (2018). High-yield production of human Dicer by transfection of human HEK293-EBNA1 cells grown in suspension. BMC Biotechnology, 18, 76.
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10

DNA-Peptide Conjugates for Liposome Fabrication

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DNA oligonucleotides were purchased from
Integrated DNA Technologies (see Supporting Information for sequences). DNA-peptide conjugates were purchased from Bio-Synthesis.
Lipids and liposome extrusion filters were purchased from Avanti Polar
Lipids. TEM grids and uranyl formate were purchased from Electron
Microscopy Sciences. JM109 E. coli cells and VCSM13
helper phage were purchased from Agilent Technologies. Ultracentrifuge
tubes were purchased from Beckman Coulter. Iodixanol was purchased
from Stemcell Technologies. Other chemicals and Amicon filters were
purchased from Millipore Sigma.
Grome M.W., Zhang Z, & Lin C. (2019). Stiffness and Membrane Anchor Density Modulate DNA-Nanospring-Induced Vesicle Tubulation. ACS Applied Materials & Interfaces, 11(26), 22987-22992.
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