Restriction enzymes and Phu DNA polymerase were purchased from New England Biolabs. pENTR/directional TOPO cloning kit (Invitrogen), pQE2 (Qiagen), were procured from the respective sources. Analytical grade chemicals and oligonucleotide primers were procured from Sigma. Malachite green phosphate assay kit (POMG-25H) was purchased from BioAssay System (Gentaur). Electron microscopy reagents were purchased from Electron Microscopy Sciences. Media components were purchased from BD Biosciences. Doxycycline hydrochloride was purchased from Biochem pharmaceutical.
Electron microscopy reagents
Manufactured by Electron Microscopy Sciences
Sourced in United States
Electron microscopy reagents are a range of chemical solutions and compounds used in the preparation and processing of samples for examination under an electron microscope. These reagents are designed to enhance the contrast, stability, and preservation of the sample, enabling high-resolution imaging and analysis of the specimen's structure and composition.
Automatically generated - may contain errors
Lab products found in correlation
Cholesterol methyl β cyclodextrin, by Merck Group (2 mentions)
Sphingomyelinase from bacillus cereus, by Merck Group (2 mentions)
Dil and bodipy tr ceramide 1 2 dihexadecanoyl sn glycero 3 phosphoethanolamine triethylammonium salt tr dhpe, by Thermo Fisher Scientific (2 mentions)
Topfluor ps, by Thermo Fisher Scientific (2 mentions)
2 hydroxypropyl β cyclodextrin, by Merck Group (2 mentions)
Methyl β cyclodextrin, by Merck Group (2 mentions)
1 2 dioleoyl sn glycero 3 phosphocholine, by Avanti Polar Lipids (2 mentions)
1 2 dioleoyl sn glycero 3 phospho l serine, by Avanti Polar Lipids (2 mentions)
1 2 distearoyl sn glycero 3 phosphoethanolamine n biotinyl polyethylene glycol 2000, by Avanti Polar Lipids (2 mentions)
Cytodex 3 microcarrier beads, by Merck Group (2 mentions)
Analytical grade chemicals, by Merck Group (1 mentions)
Pentr directional topo cloning kit, by Thermo Fisher Scientific (1 mentions)
Oligonucleotide primer, by Merck Group (1 mentions)
Restriction enzyme, by New England Biolabs (1 mentions)
Morada 1kx1k ccd camera, by Olympus (1 mentions)
Cm10 microscope, by Philips (1 mentions)
Nitrocellulose membrane, by GE Healthcare (1 mentions)
Immobilon p pvdf membrane, by Merck Group (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Sds page, by Bio-Rad (1 mentions)
8 protocols using electron microscopy reagents
1
Enzymatic Techniques for Molecular Cloning
2
Ultrastructural Analysis of Synaptic Vesicles
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Neurons were fixed with 1.2% glutaraldehyde in 0.1 M sodium cacodylate buffer, postfixed in 1% OsO4, 1.5% K4Fe(CN)6, 0.1M sodium cacodylate, en bloc stained with 0.5% uranyl magnesium acetate, dehydrated, and embedded in Embed 812. Where indicated, HRP reactions were developed with diaminobenzidene and H2O2 after the glutaraldehyde fixation step. Electron microscopy reagents were purchased from Electron Microscopy Sciences. Ultrathin sections were observed in a Philips CM10 microscope at 80 kV and images were taken with a Morada 1kx1k CCD camera (Olympus). For quantification, 20–30 pictures from each sample were used for calculating the C2-XL-HRP-labeled synaptic vesicles. 200–500 vesicles were measured for synaptic vesicle diameter analysis.
3
Comprehensive Reagent Sources for Proteomic Analyses
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Sources of materials were chemicals, Fisher Scientific (Fair Lawn, NJ, USA), or Sigma Chemical Co. (St-Louis, MO, USA); reagents and protein standards for SDS-PAGE, Bio-Rad Laboratories (Hercules, CA, USA); mass spectrometric grade chemicals and IPG strips for two-dimensional gel electrophoresis, GE Healthcare Bio-Sciences AB (Uppsala, Sweden); EDTA-free protease inhibitor tablets, Roche/Boeringher-Mannheim Biochemicals (Mannheim, Germany); nitrocellulose membrane, GE Healthcare (Piscataway, NJ, USA); PVDF Immobilon P membrane, Millipore Corp. (Billerica, MA, USA); horseradish peroxidase (HRP)-conjugated antibodies, Biosource International (Camarillo, CA, USA), Super Signal West Pico Chemiluminescent Substrate, Pierce Chemical Co. (Rockford, IL, USA); X-ray film, Phoenix Research Products (VWR, Batavia, IL, USA); electron microscopy reagents, Electron Microscopy Sciences (Hatfield, PA, USA).
4
Lipid Membrane Composition Analysis
5
Fixation and Embedding for Electron Microscopy
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Cells attached Cytodex 3 microcarrier beads (Sigma-Aldrich) were fixed in 2% glutaraldehyde-0.1M sodium cacodylate buffer pH 7.4. Cells were post-fixed with 1% OsO4 in 1.5% K4Fe(CN)6-0.1M sodium cacodylate buffer, followed by en bloc staining with 2% uranyl acetate in 50mM sodium maleate buffer pH 5.2, dehydration and embedding in Embed 812. Electron microscopy reagents were purchased from Electron Microscopy Sciences (Hatfield, PA).
6
Quantifying Clathrin-Coated Structures in Cells
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Cells were fixed in 2% glutaraldehyde-0.1 M sodium cacodylate. They were post-fixed with 1% OsO4 in 1.5% K4Fe(CN)6 and 0.1 M sodium cacodylate, en bloc stained with 0.5% uranyl magnesium acetate, dehydrated and embedded in Embed 812. Electron microscopy reagents were purchased from Electron Microscopy Sciences (Hatfield, PA). For morphometric analysis, cells whose entire perimeter was visible in the EM sections were selected and all clathrin-coated structures, including clathrin-coated vesicular profiles within 500 nm of the plasma membrane, were counted. The total number of coated structures per unit length of the plasma membrane of each cell was calculated and averages of 30 control and patient cells were obtained.
7
Lipid Membrane Composition Analysis
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BODIPY-sphingosine was a gift from Robert Bittman (Queens College of CUNY Flushing, NY). Reagents purchased from commercial sources: methyl-β-cyclodextrin, cholesterol-methyl-β-cyclodextrin (cholesterol-water soluble), 2-hydroxypropyl-β-cyclodextrin and sphingomyelinase from Bacillus cereus (Sigma-Aldrich, St. Louis, MO); Dil and BODIPY-TR-ceramide, 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt (TR-DHPE)(Life Technologies, Carlsbad, CA); TopFluor PS, brain sphingosine, egg PC, liver PE, cholesterol from sheep wool, brain PS, egg PA, brain PI(4)P, brain PI(4,5)P2, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[biotinyl(polyethylene glycol)-2000] (Avanti Polar Lipids, Alabaster, AL); and ATP, [γ-32P] (PerkinElmer, Waltham, MA). Electron microscopy reagents were purchased from Electron Microscopy Sciences.
8
Fixation and Embedding for Electron Microscopy
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