Untagged human M2 muscarinic acetylcholine receptor was expressed in Sf9 cells with the third intracellular loop replaced with T4 lysozyme, then extracted with digitonin and sodium cholate and purified by ligand affinity chromatography, then exchanged into decyl maltoside buffer. Purified receptor was crystallized by the lipidic cubic phase technique following addition of a stabilizing neopentyl glycol detergent. Data collection was performed at Advanced Photon Source beamlines 23ID-B and 23ID-D, and the structure solved by molecular replacement. Refinement statistics are given in Supplementary Table 2 .
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Glycols
Glycols
Glycols are a class of organic compounds containing two hydroxyl (-OH) groups attached to a carbon chain.
These versatile chemicals have a wide range of applications, including as solvents, antifreeze agents, and intermediates in the production of various materials.
Glycols exhibit unique physical and chemical properties, such as high boiling points, low freezing points, and the ability to form hydrogen bonds.
They play a crucial role in various industries, including pharmaceuticals, cosmetics, and automotive.
Researchers studying Glycols can leverage the power of PubCompare.ai, an AI-driven platform that enhances reproducibility and research accuracy in this field.
PubCompare.ai allows users to easily locate protocols from literature, preprints, and patents, while leveraging AI-driven comparisons to identify the best protocols and products.
Expereince the future of Glycols research optimization today with PubCompare.ai's cutting-edge technology.
These versatile chemicals have a wide range of applications, including as solvents, antifreeze agents, and intermediates in the production of various materials.
Glycols exhibit unique physical and chemical properties, such as high boiling points, low freezing points, and the ability to form hydrogen bonds.
They play a crucial role in various industries, including pharmaceuticals, cosmetics, and automotive.
Researchers studying Glycols can leverage the power of PubCompare.ai, an AI-driven platform that enhances reproducibility and research accuracy in this field.
PubCompare.ai allows users to easily locate protocols from literature, preprints, and patents, while leveraging AI-driven comparisons to identify the best protocols and products.
Expereince the future of Glycols research optimization today with PubCompare.ai's cutting-edge technology.
Most cited protocols related to «Glycols»
Buffers
Chromatography, Affinity
Cuboid Bone
Detergents
Digitonin
Glycols
Homo sapiens
Ligands
Lipids
Muramidase
Muscarinic Acetylcholine Receptor
Protoplasm
Sf9 Cells
Sodium Cholate
Buffers
Chromatography, Affinity
Cuboid Bone
Detergents
Digitonin
Glycols
Homo sapiens
Ligands
Lipids
Muramidase
Muscarinic Acetylcholine Receptor
Protoplasm
Sf9 Cells
Sodium Cholate
Human β2AR fused to an amino-terminal T4 lysozyme23 (link) was expressed and purified as described above. Following purification by alprenolol sepharose, the receptor was washed extensively with 30 μM of the low affinity antagonist atenolol while bound to FLAG affinity resin to fully displace alprenolol, then washed and eluted in buffer devoid of ligand to produce a homogeneously unliganded preparation. The receptor was then incubated for 30 minutes at room temperature with a stoichiometric excess of ligand (HBI or BI167107). A 1.3-fold molar excess of Nb6B9 was then added, and the sample was dialyzed overnight into a buffer consisting of 100 mM sodium chloride, 20 mM HEPES pH 7.5, 0.01% lauryl maltose neopentyl glycol detergent, and 0.001% cholesteryl hemisuccinate. In each case, ligand was included in the dialysis buffer at 100 nM concentration or higher. The sample was then concentrated using a 50 kDa spin concentrator and purified over a Sephadex S200 size exclusion column in the same buffer as for dialysis, and the β2AR-Nb6B9-ligand ternary complex was isolated. In the case of adrenaline, the low affinity and chemical instability of the ligand precluded overnight dialysis, so 100 μM adrenaline was added to receptor for 30 minutes at room temperature, then a 1.3-fold molar excess of Nb6B9 added and the sample was incubated for 30 minutes at room temperature. Following incubation, the sample was concentrated and immediately purified by size exclusion as above.
Following purification, samples were concentrated to A280 = 55 using a 50 kDa concentrator to minimize the detergent concentration in the final sample, then aliquoted into thin-walled PCR tubes at 8 μL per aliquot. Aliquots were flash frozen in liquid nitrogen and stored at -80 °C for crystallization trials. For crystallization, samples were thawed and reconstituted into lipidic cubic phase with a 1:1 mass:mass ratio of lipid. The lipid stock consisted of a 10:1 mix by mass of 7.7 monoacylglycerol (generously provided by Martin Caffrey) with cholesterol (Sigma). Samples were reconstituted by the two syringe mixing method10 (link) and then dispensed into glass sandwich plates using a GryphonLCP robot (Art Robbins Instruments). In the case of the β2AR-adrenaline complex, 1 mM fresh adrenaline was mixed with receptor prior to reconstitution. Crystals were grown using 30 nL protein/lipid drops with 600 nL overlay solution, which consisted of 18 – 24 % PEG400, 100 mM MES pH 6.2 to pH 6.7, and 40 – 100 mM ammonium phosphate dibasic. Crystals grew in 1 – 3 days, and were harvested and frozen in liquid nitrogen for data collection.
Following purification, samples were concentrated to A280 = 55 using a 50 kDa concentrator to minimize the detergent concentration in the final sample, then aliquoted into thin-walled PCR tubes at 8 μL per aliquot. Aliquots were flash frozen in liquid nitrogen and stored at -80 °C for crystallization trials. For crystallization, samples were thawed and reconstituted into lipidic cubic phase with a 1:1 mass:mass ratio of lipid. The lipid stock consisted of a 10:1 mix by mass of 7.7 monoacylglycerol (generously provided by Martin Caffrey) with cholesterol (Sigma). Samples were reconstituted by the two syringe mixing method10 (link) and then dispensed into glass sandwich plates using a GryphonLCP robot (Art Robbins Instruments). In the case of the β2AR-adrenaline complex, 1 mM fresh adrenaline was mixed with receptor prior to reconstitution. Crystals were grown using 30 nL protein/lipid drops with 600 nL overlay solution, which consisted of 18 – 24 % PEG400, 100 mM MES pH 6.2 to pH 6.7, and 40 – 100 mM ammonium phosphate dibasic. Crystals grew in 1 – 3 days, and were harvested and frozen in liquid nitrogen for data collection.
Alprenolol
ammonium phosphate
Atenolol
BI167107
Buffers
Cholesterol
cholesterol-hemisuccinate
Crystallization
Cuboid Bone
Detergents
Dialysis
Epinephrine
Freezing
Glycols
HEPES
Homo sapiens
Ligands
Lipids
Maltose
Molar
Monoglycerides
Nitrogen
polyethylene glycol 400
Proteins
Resins, Plant
sephadex
Sepharose
Sodium Chloride
Syringes
Apyrase
Chromatography, Affinity
Complex Mixtures
Cytokinesis
Detergents
Edetic Acid
Electron Microscopy
Enkephalin, Ala(2)-MePhe(4)-Gly(5)-
FLAG peptide
Glycols
GTP-Binding Proteins
HEPES
Homo sapiens
Maltose
manganese chloride
Micelles
Molar
Nucleotides
Peptide Hydrolases
Phosphoric Monoester Hydrolases
SDS-PAGE
Sodium Chloride
TEV protease
tris(2-carboxyethyl)phosphine
Purified DAMGO-bound μOR was mixed with a 1.2 molar excess of
Gi heterotrimer. The coupling reaction was allowed to proceed at
24 °C for 1 hour and was followed by addition of apyrase to catalyze
hydrolysis of unbound GDP, which destabilizes the nucleotide-free
complex40 . After one
more hour at 25 °C, a 4-fold volume of 20 mM Hepes pH 7.5, 100 mM NaCl,
1% lauryl maltose neopentyl glycol (L-MNG), 0.1% CHS was added
to the complexing reaction to initiate detergent exchange. After one hour
incubation at 25 °C to allow micelle exchange, 1 mM MnCl2 and
lambda phosphatase (New England Biolabs) were added to dephosphorylate the
preparation. This reaction was further incubated at 4 °C for 2 hours. To
remove excess G protein and residual DDM, the complexing mixture was purified by
M1 anti-FLAG affinity chromatography. Bound complex was first washed in a buffer
containing 1% L-MNG, followed by washes in gradually decreasing L-MNG
concentrations. The complex was then eluted in 20mM Hepes pH 7.5, 100mM NaCl,
0.01% MNG/0.001% CHS, 300 nM DAMGO, 5 mM EDTA, and FLAG peptide.
The eluted complex was supplemented with 100 μM TCEP to provide a
reducing environment. The tobacco etch virus (TEV) protease and human rhinovirus
3C protease were added to cleave the flexible μOR amino- and carboxy-
termini. Finally, a 1.2 molar excess of scFv16 was added to the preparation.
Once cleavage of the termini was confirmed by SDS-PAGE, the
μOR-Gi-scFv16 complex was purified by size exclusion
chromatography on a Superdex 200 10/300 column in 20mM Hepes pH 7.5, 100mM NaCl,
300 nM DAMGO, 0.00075% MNG and 000025% GDN. Peak fractions were
concentrated to ~7 mg/mL for electron microscopy studies.
Gi heterotrimer. The coupling reaction was allowed to proceed at
24 °C for 1 hour and was followed by addition of apyrase to catalyze
hydrolysis of unbound GDP, which destabilizes the nucleotide-free
complex40 . After one
more hour at 25 °C, a 4-fold volume of 20 mM Hepes pH 7.5, 100 mM NaCl,
1% lauryl maltose neopentyl glycol (L-MNG), 0.1% CHS was added
to the complexing reaction to initiate detergent exchange. After one hour
incubation at 25 °C to allow micelle exchange, 1 mM MnCl2 and
lambda phosphatase (New England Biolabs) were added to dephosphorylate the
preparation. This reaction was further incubated at 4 °C for 2 hours. To
remove excess G protein and residual DDM, the complexing mixture was purified by
M1 anti-FLAG affinity chromatography. Bound complex was first washed in a buffer
containing 1% L-MNG, followed by washes in gradually decreasing L-MNG
concentrations. The complex was then eluted in 20mM Hepes pH 7.5, 100mM NaCl,
0.01% MNG/0.001% CHS, 300 nM DAMGO, 5 mM EDTA, and FLAG peptide.
The eluted complex was supplemented with 100 μM TCEP to provide a
reducing environment. The tobacco etch virus (TEV) protease and human rhinovirus
3C protease were added to cleave the flexible μOR amino- and carboxy-
termini. Finally, a 1.2 molar excess of scFv16 was added to the preparation.
Once cleavage of the termini was confirmed by SDS-PAGE, the
μOR-Gi-scFv16 complex was purified by size exclusion
chromatography on a Superdex 200 10/300 column in 20mM Hepes pH 7.5, 100mM NaCl,
300 nM DAMGO, 0.00075% MNG and 000025% GDN. Peak fractions were
concentrated to ~7 mg/mL for electron microscopy studies.
Apyrase
Chromatography, Affinity
Complex Mixtures
Cytokinesis
Detergents
Edetic Acid
Electron Microscopy
Enkephalin, Ala(2)-MePhe(4)-Gly(5)-
FLAG peptide
Glycols
GTP-Binding Proteins
HEPES
Homo sapiens
Maltose
manganese chloride
Micelles
Molar
Nucleotides
Peptide Hydrolases
Phosphoric Monoester Hydrolases
SDS-PAGE
Sodium Chloride
TEV protease
tris(2-carboxyethyl)phosphine
Most recents protocols related to «Glycols»
Example 1
A glass article comprising aluminosilicate glass as commercially available under the designation AS 87 was first cleaned in an industrial dishwasher in order to clean the surfaces of the glass article from dirt such as grease, fingerprints, or the like. Then, a 0.25 wt % solution of a modified fluoroalkyl oligosiloxane in ethanol (1 g of fluoroalkyl oligosiloxane in 395 g of ethanol) was applied over the entire surface of the glass article. In this way, a surface of the glass article was made hydrophobic. More generally, without being limited to the example specifically described here, it is also possible that only an area of the at least one surface of the glass article is made hydrophobic, for example by covering the area or areas of the at least one surface, which are not intended to be hydrophobized. Subsequently, a 5% solution of a surfactant in ethanol and a mixture of different glycol ethers was applied to the hydrophobized surface of the glass article by spraying. Then, plasma etching was performed in a CF4-containing atmosphere in a microwave plasma at a pressure of 10 mbar.
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aluminosilicate
Atmosphere
Ethanol
Ethers
Glare
Glycols
Microwaves
Plasma
Pressure
Surface-Active Agents
Example 2
The Bioceramic compositions in Table 2, below, were prepared by mixing the liquid component (carrier) with the solid components in a mechanical stirrer, in the following sequence: sorosilicate, radiopacifier, rheology control agent and setting agent with speed below 500 rpm, approximately 45 minutes until complete homogenization.
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akermanite
Calcium, Dietary
Glycols
Oxides
Paste
Polyethylenes
Potassium-37
Silicon-29
Strontium
Sulfates, Inorganic
tungstate
Zirconium
Example 2
A glass article comprising a lithium aluminum silicate glass as commercially available under the designation LAS 80 was first cleaned in an industrial dishwasher in order to clean the surfaces of the glass article from dirt such as grease, fingerprints, or the like. Then, a 0.25 wt % solution of a modified fluoroalkyl oligosiloxane in ethanol (1 g of fluoroalkyl oligosiloxane in 395 g of ethanol) was applied over the entire surface of the glass article. In this way, a surface of the glass article was made hydrophobic. Subsequently, a 5% solution of a surfactant in ethanol and a mixture of different glycol ethers was applied to the hydrophobized surface of the glass article by spraying. Then, plasma etching was performed in a CF4-containing atmosphere in a microwave plasma at a pressure of 10 mbar.
Again, it is generally possible here that the at least one surface is not hydrophobized over its entire surface area, but only in an area of the surface.
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Aluminum
Atmosphere
Ethanol
Ethers
Glare
Glycols
Lithium
Microwaves
Plasma
Pressure
Silicates
Surface-Active Agents
Example 14
A 1 L round bottom flask equipped with a condensor was placed under a N2 atmosphere and charged with propylene imine (80.0 gram), n-butyl glycidyl ether (126.0 gram) and K2CO3 (10.00 gram) and heated to 80° C. in 30 min, after which the mixture was stirred for 21 h at T=80° C. After filtration the excess of PI was removed in vacuo, followed by further purification via vacuum distillation, resulting in a colorless low viscous liquid.
1.92 grams of the resulting material (1-butoxy-3-(2-methylaziridin-1-yl)propan-2-ol) was charged to a reaction flask equipped with a thermometer, together with 0.02 grams of bismuth neodecanoate and 19 grams of dimethylformamide. The mixture was stirred with a mechanical upper stirrer under a nitrogen atmosphere and heated to 50° C. A solution of 2.00 grams of Desmodur N 3600 in 19 grams of dimethylformamide was then added dropwise in 45 minutes to the reaction flask, whereafter the mixture was heated further to 70° C. Samples were taken at regular intervals and the reaction progress was monitored using a Bruker Alpha FT-IR spectrometer until no NCO-stretch at 2200-2300 cm−1 was observed. The solvent was removed in vacuo to obtain a clear, yellowish highly viscous liquid. The calculated molecular weight of the theoretical main component was 1065.74 Da, chemical structure is shown below.
Molecular weight was confirmed by Maldi-TOF-MS: Calcd. [M+Na+]=1088.74 Da; Obs. [M+Na+]=1088.76 Da. The following components with a mass below 580 Da were determined by LC-MS and quantified:
Performance of the synthesized compound as a crosslinker was assessed using spot tests on coating surfaces, based on procedures from the DIN 68861-1 standard. For these tests, 0.58 parts of the composition were mixed with 0.60 parts of Proglyde™ DMM (dipropylene glycol dimethyl ether, mixture of isomers) and incubated at 80° C. for 10 minutes under regular agitation. Subsequently, 0.79 parts of the resulting solution were added to 20 parts of NeoRez® R-1005 under continuous stirring, and the resulting mixture was further stirred for 30 minutes. Afterwards, this coating composition was filtered and applied to Leneta test cards using 100 μm wire rod applicators (Test 14-1). For reference, films were also cast from the same composition lacking a crosslinker (Test 14-2). The films were dried for 16 hours at 25° C., then annealed at 50° C. for 1 hour and further dried for 24 hours at 25° C. Subsequently, a piece of cotton wool was soaked in 1:1 EtOH:demineralized water and placed on the film for various timespans. After removal of the EtOH and 60 minutes recovery, the following results were obtained (a score of 1 indicates complete degradation of the film, 10 indicates no damage visible):
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Atmosphere
Aziridines
Bismuth
CD3EAP protein, human
Desmodur N
dimethyl ether
Dimethylformamide
Distillation
Ethanol
farnesyl-protein transferase-alpha
Filtration
Glycols
Gossypium
Isomerism
Liver Extracts
Mutagenicity Tests
n-butyl glycidyl ether
Nitrogen
potassium carbonate
propyleneimine
Solvents
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Thermometers
Vacuum
Viscosity
Example 1
A formulation of a suspension composition of the type in the present disclosure for 1000 gram fluid is listed in Table 1 below. The suspension composition was prepared and used in Examples 1 and 2.
The suspension composition was firstly used in stability tests. The suspension composition was kept static in a standing 25 ml measuring cylinder to observe mixture stability.
After 21 days from preparation, density of the suspension composition was checked from top, middle and bottom portion of the suspension composition and shown in Table 2.
As shown in
Physical properties were measured for the suspension composition and shown in Table 3 below.
Example 2
Wellbore servicing fluids were prepared using a dry powder suspending agent or the suspension composition in Example 1. Test conditions and formulas of the wellbore servicing fluids are listed in Tables 4 and 5. The amounts of the cement blend composition are based on the total weight of the cement blend. The amount of the dry powder suspending agent is based on the total weight of the cement blend, while the dry powder suspending agent is not a part of the cement blend. Both of the wellbore servicing fluids had a density of 14.60 lbm/gal and a specific gravity of 1.75. The amount of the dry powder suspending agent in wellbore servicing fluid 1 (WSF1) was 1.3 g per 600 ml WSF1, which was equivalent to the amount of the crosslinked guar gum in wellbore servicing fluid 2 (WSF2).
Table 6 below shows 24 hr sonic compressive strength is lower in WSF2 compared to WSF1, however other properties are comparable.
Table 7 shows that the rheology data measured by a Fann® Model 35 viscometer for WSF 1 and WSF 2 are comparable.
Further, WSF1 and WSF2 were cured at 168° F./5.000 psig for 7 days and then tested for mechanical properties. The results are in Table 8 below.
The experiments demonstrate the following. 7 days curing data shows there was no adverse effect of the use of the suspension composition on mechanical properties of set cement. UCA Compressive Strength shows a slight delay in strength development for WSF2. Regarding to other slurry properties such as mixability, free fluid, rheology, gel strength, and fluid loss, there was no adverse effect of the use of the suspension composition by comparing WSF1 and WSF2.
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Dental Cements
Diet, Formula
GAL-1
Glycols
Gravity
guar gum
LGALS9 protein, human
Physical Processes
Powder
Pressure
Viscosity
Top products related to «Glycols»
Sourced in United States
Lauryl maltose neopentyl glycol is a non-ionic detergent used in the solubilization and stabilization of membrane proteins. It is a milder detergent compared to traditional ionic detergents, making it suitable for maintaining the native structure and function of proteins.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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Polyvinyl alcohol is a synthetic, water-soluble polymer. It is commonly used as a raw material in the production of various laboratory equipment and supplies.
Sourced in United States, Germany, Netherlands, United Kingdom, Czechia, Israel
The Vitrobot Mark IV is a cryo-electron microscopy sample preparation instrument designed to produce high-quality vitrified specimens for analysis. It automates the process of blotting and plunge-freezing samples in liquid ethane, ensuring consistent and reproducible sample preparation.
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Acetone is a colorless, volatile, and flammable liquid. It is a common solvent used in various industrial and laboratory applications. Acetone has a high solvency power, making it useful for dissolving a wide range of organic compounds.
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
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Tween 80 is a non-ionic surfactant and emulsifier. It is a viscous, yellow liquid that is commonly used in laboratory settings to solubilize and stabilize various compounds and formulations.
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1,4-butanediol is a colorless, viscous chemical compound that is commonly used as a laboratory reagent. It has a molecular formula of C4H10O2 and a molecular weight of 90.12 g/mol. 1,4-butanediol is a versatile compound that can be used for various applications in research and development.
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Ethanol is a clear, colorless liquid chemical compound commonly used in laboratory settings. It is a key component in various scientific applications, serving as a solvent, disinfectant, and fuel source. Ethanol has a molecular formula of C2H6O and a range of industrial and research uses.
More about "Glycols"
Glycols are a diverse class of organic compounds characterized by the presence of two hydroxyl (-OH) groups attached to a carbon chain.
These versatile chemicals exhibit a wide range of applications, serving as solvents, antifreeze agents, and intermediates in the production of various materials.
Glycols, such as ethylene glycol, propylene glycol, and 1,4-butanediol, possess unique physical and chemical properties, including high boiling points, low freezing points, and the ability to form hydrogen bonds.
These properties make them indispensable in numerous industries, including pharmaceuticals, cosmetics, and automotive.
Researchers studying glycols can leverage the power of PubCompare.ai, an AI-driven platform that enhances reproducibility and research accuracy in this field.
PubCompare.ai allows users to easily locate protocols from literature, preprints, and patents, while utilizing AI-driven comparisons to identify the best protocols and products.
This cutting-edge technology can help optimize glycols research, including the use of related compounds like lauryl maltose neopentyl glycol, FBS, polyvinyl alcohol, Vitrobot Mark IV, acetone, penicillin/streptomycin, Tween 80, and ethanol.
Experience the future of glycols research optimization today with PubCompare.ai's innovative solutions.
These versatile chemicals exhibit a wide range of applications, serving as solvents, antifreeze agents, and intermediates in the production of various materials.
Glycols, such as ethylene glycol, propylene glycol, and 1,4-butanediol, possess unique physical and chemical properties, including high boiling points, low freezing points, and the ability to form hydrogen bonds.
These properties make them indispensable in numerous industries, including pharmaceuticals, cosmetics, and automotive.
Researchers studying glycols can leverage the power of PubCompare.ai, an AI-driven platform that enhances reproducibility and research accuracy in this field.
PubCompare.ai allows users to easily locate protocols from literature, preprints, and patents, while utilizing AI-driven comparisons to identify the best protocols and products.
This cutting-edge technology can help optimize glycols research, including the use of related compounds like lauryl maltose neopentyl glycol, FBS, polyvinyl alcohol, Vitrobot Mark IV, acetone, penicillin/streptomycin, Tween 80, and ethanol.
Experience the future of glycols research optimization today with PubCompare.ai's innovative solutions.