Simulations using Gromacs 4.5.3 were carried out for the following four folded globular proteins: bovine pancreatic trypsin inhibitor (BPTI), ubiquitin, GB3 and hen Lysozyme, starting from the published experimental structures (PDB entries 5PTI48 (link), 1UBQ49 (link), 1P7E50 (link), 6LYT51 respectively). Each protein was solvated in a truncated octahedron simulation cell filled with TIP3P water, with nearest distance between images of 45 Å for all proteins except for lysozyme, for which the distance was 60 Å. Sodium and chloride ions were added as needed to yield a final salt concentrations of ~100 mM, with adjustments to ensure charge neutrality. For each protein, a 100-step SD energy minimization of the whole system was performed, followed by 200 ps of MD at a constant pressure of 1 bar and temperature of 300 K, in which harmonic positional restraints of 2.39 kcal/mol/Å2 were applied to each Cartesian component of each protein non-hydrogen atom using the minimized structure as a reference. Each protein was then simulated at a constant pressure of 1 bar and a temperature of 300 K for 200 ns. Pressure was regulated by a Parinello-Rahman barostat52 with a coupling time of 2.5 ps; otherwise all details were as described for Ac-(AAQAA)3-NH2 above.
>
Chemicals & Drugs
>
Organic Chemical
>
A 300
A 300
A 300: A comprehensive, AI-powered platform that assists researchers in optimizing their research protocols for reproducibility and accuracy.
Discover how PubCompare.ai enables you to easily locate, compare, and identify the best protocols and products for your 300 research projects.
Experiance streamlined, data-driven decision making through intelligent protocol comparisons from literature, preprints, and patents.
Simplify your research workflow and ensure robust, reliable results with this innovative tool.
Discover how PubCompare.ai enables you to easily locate, compare, and identify the best protocols and products for your 300 research projects.
Experiance streamlined, data-driven decision making through intelligent protocol comparisons from literature, preprints, and patents.
Simplify your research workflow and ensure robust, reliable results with this innovative tool.
Most cited protocols related to «A 300»
A 300
Aprotinin
Cells
Chlorides
Eye
hen egg lysozyme
Hydrogen
Ions
Muramidase
Pressure
Proteins
Sodium
Sodium Chloride
Ubiquitin
A 300
ECHO protocol
Head
Human Body
Neoplasm Metastasis
Sepharose
Vibration
All MRI and MRF data were acquired on a 1.5T whole body scanner (Siemens Espree, Siemens Healthcare, Erlangen, Germany) with a 32 channel head receiver coil (Siemens Healthcare, Erlangen, Germany). A square field of view of 300 × 300 mm2 was covered with a matrix of 128×128 pixels. The slice thickness was 5 mm. Images from each acquisition block were reconstructed separately using non-uniform Fourier transform (NUFFT)50 . The resultant time series of images was used to determine the value for the parameters (T1, T2, M0 and off-resonance) as described above.
In vivo experiments were performed with IRB guidelines, including written informed consent. For the fully sampled spiral acquisition shown inSupplementary Movie1 , 48 repetitions were acquired, each with a different interleaf of the total acquisition. A recovery time of 5 seconds was used in between various acquisitions and this was taken into account in the simulated dictionary.
For the phantom study shown inFigure 2 and 4 , eight cylindrical phantoms were constructed with varying concentrations of GdCl3 (Aldrich) and agarose (Sigma) to yield different T1 and T2 values ranging from 67 to 1700 ms and 30 to 200 ms, respectively. Standard Spin Echo (SE) sequences were used to quantify T1 and T2 separately (T1 quantification: 13 TRs ranging from 50 to 5000 ms, TE = 8.5 ms, total acquisition time = 33.4 minutes; T2 quantification: Spin Echo sequences with TEs = [15 30 45 60 90 150 200 300 400] ms, TR = 10000 ms, total acquisition time = 3.2 hours.). T1 values were calculated pixel-wise using a standard three-parameter nonlinear least squares fitting routine to solve the equation: S(TR) = a + beT R/T1. T2 values were determined in a pixel-wise fashion using a two-parameter nonlinear least squares fitting routine to solve the equation S(TE) = ae−T E/T2, DESPOT1 and DESPOT2 sequences using a fully sampled spiral readout were implemented based on the acquisition values from Deoni et al30 (link): DESPOT1: FA: 4° and 15°, TR:13.6 ms, DESPOT2: FA: 15° and 55°, TR=10.8 ms. The T1 and T2 values were calculated from the equations provided by Deoni et al 30 (link). A 20 s waiting period was used in between the different acquisitions. The initial 10 s of data acquisition was not used in order to ensure that the signal was in steady-state for each of the DESPOT acquisitions. In the following analysis of efficiency, only the pure time of data acquisition for the steady-state DESPOT images is used. For DESPOT1 this was 1.27 s and for DESPOT2 it was 2.29 s (which includes the time for the required DESPOT1 acquisition.)
In vivo experiments were performed with IRB guidelines, including written informed consent. For the fully sampled spiral acquisition shown in
For the phantom study shown in
A 300
ECHO protocol
Head
Human Body
Neoplasm Metastasis
Sepharose
Vibration
The gene symbol and sequence for each candidate reference gene was retrieved from the affymetrix microarray probe set and compared to the NCBI E. coli BL21 (DE3) genomic sequence (NC_012971.1). Vector NTI Advance 10 (Invitrogen) was used to design two sets of primers for each target gene. The first set of primers generated amplicons of ~300 bp which were used as standard templates for qPCR of the targeted gene. The second set of primers was used for qPCR assays and was designed to amplify a ~100 bp region within each ~300 bp template. All products generated after amplifications were verified by gel-electrophoresis.
Full text: Click here
A 300
Biological Assay
BP 100
Cloning Vectors
Electrophoresis
Escherichia coli
Genes
Genome
Microarray Analysis
Oligonucleotide Primers
All MD simulations were
performed with NAMD40 (link) employing CHARMM-formatted
parameter files41 (link) for all force fields
tested, which are provided in theSupporting Information . For all simulations, a temperature of 300 K and pressure of 1 atm
were maintained with a Nose–Hoover Langevin piston barostat
with a piston period of 100 fs and a piston dampening time scale of
50 fs and a Langevin thermostat with a damping coefficient of 1 ps–1. Nonbonded cutoffs were employed at 11 Å with
a smoothing function starting at 9 Å, with particle mesh Ewald
used to treat long-range electrostatics. The systems were solvated
in cubic water boxes with edge lengths ranging from 25 to 58 Å.
Sodium and chloride ions were added to neutralize the charges in the
system and provide approximately a 150 mM concentration of salt. A
2 fs time step was employed with the use of SHAKE and SETTLE.
Triplicate 205 ns simulations were run for an unblocked alanine pentapeptide
(Ala5) with and glycine tripeptide (Gly3) with
protonated C-termini with the first 5 ns discarded as equilibration.
The remaining amino acids, with the exception of proline, were simulated
for 205 ns as blocked dipeptides, again in triplicate with the first
5 ns discarded as equilibration. Values and error bars throughout
the paper represent the mean and standard deviation of the calculated
quantities from the triplicate runs. Ala5 and Gly3 simulations were run with each of the four weighting temperatures
examined in this work, as well as the previous OPLS-AA and OPLS-AA/L
force field. Dipeptide simulations were performed with OPLS-AA, OPLS-AA/L,
and the new parameters optimized at 2000 K. As each system was studied
for 600 ns with at least three different force fields, over 50 μs
of validating simulations have been executed. In analyzing the molecular
dynamics simulations for the short alanine and glycine peptides, the
definitions of secondary structure, the three sets of Karplus parameters
for calculating J couplings, and the experimental
error values used to calculate χ2 from Best et al.42 (link) were employed. For the dipeptide simulations,
only the first set of Karplus parameters, that of Hu and Bax,43 was employed. χ1 rotamer populations
were determined by dividing the range of χ1 values
into three equal sized bins, corresponding to the p (+60°), t
(180°) and m (−60°) conformers. Definitions of p,
t, and m for valine, isoleucine, and threonine were adopted from the
work of Dunbrak and co-workers27 (link) and are
depicted in Figure1 .
The proteins ubiquitin and GB3 were started from the PDB
structures 1UBQ(44 (link)) and 1P7E(45 (link)) and gradually
heated to 300 K over
400 ps before 205 ns simulations were run. Both the heating period
and the first 5 ns were discarded as equilibration, and simulations
were performed in triplicate for each protein. All other simulation
parameters were identical to those used for the dipeptides. For calculation
of backbone J couplings of the full protein, both
the 1997 empirical Karplus parameters43 used for the dipeptides and another empirical model developed from
work with GB346 (link) are employed. Side chain J couplings were calculated for couplings to methyl side
chains with the set of Karplus parameters developed by Vögeli
et al.,46 (link) while all other couplings employed
Karplus parameters from Perez et al.48 (link)
performed with NAMD40 (link) employing CHARMM-formatted
parameter files41 (link) for all force fields
tested, which are provided in the
were maintained with a Nose–Hoover Langevin piston barostat
with a piston period of 100 fs and a piston dampening time scale of
50 fs and a Langevin thermostat with a damping coefficient of 1 ps–1. Nonbonded cutoffs were employed at 11 Å with
a smoothing function starting at 9 Å, with particle mesh Ewald
used to treat long-range electrostatics. The systems were solvated
in cubic water boxes with edge lengths ranging from 25 to 58 Å.
Sodium and chloride ions were added to neutralize the charges in the
system and provide approximately a 150 mM concentration of salt. A
2 fs time step was employed with the use of SHAKE and SETTLE.
Triplicate 205 ns simulations were run for an unblocked alanine pentapeptide
(Ala5) with and glycine tripeptide (Gly3) with
protonated C-termini with the first 5 ns discarded as equilibration.
The remaining amino acids, with the exception of proline, were simulated
for 205 ns as blocked dipeptides, again in triplicate with the first
5 ns discarded as equilibration. Values and error bars throughout
the paper represent the mean and standard deviation of the calculated
quantities from the triplicate runs. Ala5 and Gly3 simulations were run with each of the four weighting temperatures
examined in this work, as well as the previous OPLS-AA and OPLS-AA/L
force field. Dipeptide simulations were performed with OPLS-AA, OPLS-AA/L,
and the new parameters optimized at 2000 K. As each system was studied
for 600 ns with at least three different force fields, over 50 μs
of validating simulations have been executed. In analyzing the molecular
dynamics simulations for the short alanine and glycine peptides, the
definitions of secondary structure, the three sets of Karplus parameters
for calculating J couplings, and the experimental
error values used to calculate χ2 from Best et al.42 (link) were employed. For the dipeptide simulations,
only the first set of Karplus parameters, that of Hu and Bax,43 was employed. χ1 rotamer populations
were determined by dividing the range of χ1 values
into three equal sized bins, corresponding to the p (+60°), t
(180°) and m (−60°) conformers. Definitions of p,
t, and m for valine, isoleucine, and threonine were adopted from the
work of Dunbrak and co-workers27 (link) and are
depicted in Figure
The proteins ubiquitin and GB3 were started from the PDB
structures 1UBQ(44 (link)) and 1P7E(45 (link)) and gradually
heated to 300 K over
400 ps before 205 ns simulations were run. Both the heating period
and the first 5 ns were discarded as equilibration, and simulations
were performed in triplicate for each protein. All other simulation
parameters were identical to those used for the dipeptides. For calculation
of backbone J couplings of the full protein, both
the 1997 empirical Karplus parameters43 used for the dipeptides and another empirical model developed from
work with GB346 (link) are employed. Side chain J couplings were calculated for couplings to methyl side
chains with the set of Karplus parameters developed by Vögeli
et al.,46 (link) while all other couplings employed
Karplus parameters from Perez et al.48 (link)
A-A-1 antibiotic
A 300
Alanine
Amino Acids
Chlorides
Cuboid Bone
Dipeptides
Electrostatics
Glycine
Ions
Isoleucine
Nose
Peptides
Pressure
Proline
Proteins
Sodium
Sodium Chloride
Threonine
Tremor
Ubiquitin
Valine
Vertebral Column
Most recents protocols related to «A 300»
Example 3
Moulded Silicone Pressure Sensitive Adhesive Body:
Dow Corning 7-9800 A&B (mixing ration between A and Bis 1:1 by weight) were used for production of a PDMS based adhesive body. A mould having a triangular shape (each side of the triangular mould having a distance of 300 mm, the center part having a thickness of 0.5 mm and the edge having a thickness of 0.1 mm) was used. The components were thoroughly mixed and applied on a 50 μm cover layer of silicone rubber lining in the female part of a triangular mould and a male mould part was placed on top, said part lined with a low density polyethylene release liner. The adhesive was cured in an oven at 100 degree C. for 15 minutes. After curing the adhesive was punched out of the mould and a dent in the centre of the adhesive body device for embedment of an electronic sensing system was punched out.
Full text: Click here
A 300
Dental Cavity Liner
Females
Fungus, Filamentous
Human Body
Males
Polyethylene, Low-Density
Pressure
Silicone Elastomers
Silicones
Example 1
<Step (A): Synthesis of porous particle having glycidyl group>
27.8 g of glycidyl methacrylate (trade name: Blemmer G (registered trademark) manufactured by NOF Corporation), 11.3 g of glycerin-1,3-dimethacrylate (trade name: NK Ester 701, SHIN-NAKAMURA CHEMICAL Co., Ltd.), and 1.9 g of 2,2′-azobis(2,4-dimethylvaleronitrile) were dissolved in 58.7 g of diethyl succinate as a diluent, and nitrogen gas was bubbled for 30 minutes to provide an oil phase.
Next, separately from the oil phase, 10.0 g of PVA-224 (manufactured by Kuraray Co., Ltd., polyvinyl alcohol having a degree of saponification of 87.0% to 89.0%) as a dispersion stabilizer and 10.0 g of sodium chloride as a salting-out agent were dissolved in 480 g of ion exchanged water to provide an aqueous phase.
The aqueous phase and the oil phase were placed in a separable flask and dispersed at a rotation speed of 430 rpm for 20 minutes using a stirring rod equipped with a half-moon stirring blade, then the inside of the reactor was purged with nitrogen, and the reaction was carried out at 60° C. for 16 hours.
After that, the resulting polymer was transferred onto a glass filter and thoroughly washed with hot water at about 50 to 80° C., denatured alcohol, and water in the order presented to obtain 100.4 g of a porous particle (carrier al).
The amount of glycidyl methacrylate used was 79.8 mol % based on the total amount of the monomers, and the amount of glycerin-1,3-dimethacrylate used was 20.2 mol % based on the total amount of the monomers.
<Step (B): Introduction reaction of alkylene group>
98 g of the carrier α1 was weighed onto a glass filter and thoroughly cleaned with diethylene glycol dimethyl ether. After cleaning, the carrier α1 was placed in a 1 L separable flask, 150 g of diethylene glycol dimethyl ether and 150 g (920 mol % based on glycidyl methacrylate) of 1,4-butanediol were placed in the separable flask, and stirring and dispersion were carried out.
After that, 1.5 ml of a boron trifluoride diethyl ether complex was added, the temperature was raised to 80° C. while stirring at 200 rpm, and the resulting mixture was subjected to the reaction for 4 hours.
The mixture was cooled, then the porous particle (carrier β1) bonded to a diol compound including an alkylene group in the structure thereof was collected by filtration and then washed with 1 L of ion exchanged water to obtain 152 g of a carrier β1.
The progress of the reaction was confirmed by the following procedure.
A part of the dry porous particle into which an alkylene group had been introduced was mixed with potassium bromide, and the resulting mixture was pelletized by applying a pressure and then measured using FT-IR (trade name: Nicolet (registered trademark) iS10, manufactured by Thermo Fisher Scientific Inc.) to check the height of an absorbance peak at 908 cm−1 due to the glycidyl group in the infrared absorption spectrum.
As a result, no absorbance peak at 908 cm−1 was observed by FT-IR.
<Step (C): Introduction Reaction of Glycidyl Group>
150 g of the carrier β1 was weighed onto a glass filter and thoroughly cleaned with dimethylsulfoxide.
After cleaning, the carrier β1 was placed in a separable flask, 262.5 g of dimethyl sulfoxide and 150 g of epichlorohydrin were added, the resulting mixture was stirred at room temperature, 37.5 ml of a 30% sodium hydroxide aqueous solution (manufactured by KANTO CHEMICAL CO., INC.) was further added, and the resulting mixture was heated to 30° C. and stirred for 6 hours.
After completion of the reaction, the obtained product was transferred onto a glass filter and thoroughly washed with water, acetone, and water in the order presented to obtain 172 g of a porous particle into which a glycidyl group had been introduced (carrier γ1).
The introduction density of the glycidyl group in the obtained carrier γ1 was measured by the following procedure.
5.0 g of the carrier γ1 was sampled, and the dry mass thereof was measured and as a result, found to be 1.47 g. Next, the same amount of the carrier γ1 was weighed into a separable flask and dispersed in 40 g of water, 16 mL of diethylamine was added while stirring at room temperature, and the resulting mixture was heated to 50° C. and stirred for 4 hours. After completion of the reaction, the reaction product was transferred onto a glass filter and thoroughly washed with water to obtain a porous particle A into which diethylamine had been introduced.
The obtained porous particle A was transferred into a beaker and dispersed in 150 mL of a 0.5 mol/L potassium chloride aqueous solution, and titration was carried out using 0.1 mol/L hydrochloric acid with the point at which the pH reached 4.0 as the neutralization point.
From this, the amount of diethylamine introduced into the porous particle A into which diethylamine had been introduced was calculated, and the density of the glycidyl group of the carrier γ1 was calculated from the following expression.
As a result, the density of the glycidyl group was 880 μmol/g.
Density(μmol/g) of glycidyl group={0.1×volume(μL) of hydrochloric acid at neutralization point/dry mass(g) of porous particle into which glycidyl group has been introduced}<Step (D): Introduction Reaction of Polyol>
150 g of the carrier γ1, 600 mL of water, and 1000 g (13000 mol % based on glycidyl group) of D-sorbitol (log P=−2.20, manufactured by KANTO CHEMICAL CO., INC.) were placed in a 3 L separable flask and stirred to form a dispersion.
After that, 10 g of potassium hydroxide was added, the temperature was raised to 60° C. while stirring at 200 rpm, and the resulting mixture was subjected to the reaction for 15 hours.
The mixture was cooled, and then the reaction product was collected by filtration and washed thoroughly with water to obtain 152 g of a porous particle into which polyol had been introduced (carrier 61).
The obtained carrier 61 was classified into 16 to 37 μm using a sieve to obtain 140.5 g of a packing material 1.
<Evaluation of Alkali Resistance>
The alkali resistance was evaluated by calculating the amount of a carboxy group produced by hydrolysis of sodium hydroxide according to the following procedure.
First, 4 g of the packing material was dispersed in 150 mL of a 0.5 mol/L potassium chloride aqueous solution, and titration was carried out using 0.1 mol/L sodium hydroxide aqueous solution with the point at which the pH reached 7.0 as the neutralization point. From this, the amount of a carboxy group before hydrolysis included in the packing material was calculated from the following expression.
Amount(μmol/mL) of carboxy group=0.1×volume(μL) of sodium hydroxide aqueous solution at the time of neutralization/apparent volume (mL) of packing material
Here, the apparent volume of the packing material is the volume of the packing material phase measured after preparing a slurry liquid by dispersing 4 g of the packing material in water, transferring the slurry liquid to a graduated cylinder, and then allowing the same to stand for a sufficient time.
Subsequently, 4 g of the packing material was weighed into a separable flask, 20 mL of a 5 mol/L sodium hydroxide aqueous solution was added, and the resulting mixture was treated at 50° C. for 20 hours while stirring at 200 rpm. The mixture was cooled, then the packing material was collected by filtration, then washed with a 0.1 mol/L HCl aqueous solution and water in the order presented, and the amount of a carboxy group contained in the obtained packing material was calculated by the same method as above. From the difference between the amount of a carboxy group before and that after the reaction with the 5 mol/L sodium hydroxide aqueous solution, the amount of a carboxy group produced by the reaction with the 5 mol/L sodium hydroxide aqueous solution was calculated. As a result, the amount of a carboxy group produced was 21 μmol/mL.
If the amount of a carboxy group produced is 40 μmol/mL or less, the alkali resistance is considered to be high.
<Evaluation of Non-Specific Adsorption>
The obtained packing material was packed into a stainless steel column (manufactured by Sugiyama Shoji Co., Ltd.) having an inner diameter of 8 mm and a length of 300 mm by a balanced slurry method. Using the obtained column, a non-specific adsorption test was carried out by the method shown below.
The column packed with the packing material was connected to a Shimadzu Corporation HPLC system (liquid feed pump (trade name: LC-10AT, manufactured by Shimadzu Corporation), autosampler (trade name: SIL-10AF, manufactured by Shimadzu Corporation), and photodiode array detector (trade name: SPD-M10A, manufactured by Shimadzu Corporation)), and a 50 mmol/L sodium phosphate buffer aqueous solution as a mobile phase was passed at a flow rate of 0.6 mL/min.
Using the same sodium phosphate aqueous solution as the mobile phase as a solvent, their respective sample solutions of 0.7 mg/mL thyroglobulin (Mw of 6.7×105), 0.6 mg/mL γ-globulin (Mw of 1.6×105), 0.96 mg/mL BSA (Mw of 6.65×104), 0.7 mg/mL ribonuclease (Mw of 1.3×104), 0.4 mg/mL aprotinin (Mw of 6.5×103), and 0.02 mg/mL uridine (Mw of 244) (all manufactured by Merck Sigma-Aldrich) are prepared, and 10 μL of each is injected from the autosampler.
The elution time of each observed using the photodiode array detector at a wavelength of 280 nm was compared to confirm that there was no contradiction between the order of elution volume and the order of molecular weight size.
As a result, the elution volumes of the samples from the column packed with the packing material 1 were 8.713 mL, 9.691 mL, 9.743 mL, 10.396 mL, 11.053 mL, and 11.645 mL, and it was confirmed that there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that no non-specific adsorption was induced. When there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof, there was no non-specific adsorption, which is indicated as 0 in Table 1, and when there was a contradiction therebetween, non-specific adsorption was induced, which is thus indicated as X.
The porous particle (carrier al) obtained in the same manner as in Example 1 was subjected to the step D of Example 1.
<Step (D): Introduction Reaction of Polyol>
98 g of carrier al, 600 mL of water, and 1000 g (3050 mol % based on glycidyl group) of D-sorbitol (manufactured by KANTO CHEMICAL CO., INC.) were placed in a 3 L separable flask and stirred to form a dispersion.
After that, 10 g of potassium hydroxide was added, the temperature was raised to 60° C. while stirring at 200 rpm, and the resulting mixture was subjected to the reaction for 15 hours.
The mixture was cooled, and then the reaction product was collected by filtration and washed thoroughly with water to obtain 130 g of a porous particle into which a polyol had been introduced (carrier δ7).
The carrier δ7 was classified into 16 to 37 μm using a sieve to obtain 115 g of a packing material 7.
The alkali resistance of the obtained packing material 7 was evaluated in the same manner as in Example 1. As a result, the amount of a carboxy group produced in the packing material 7 was 120.3 μmol/mL, resulting in poor alkali resistance.
Further, the non-specific adsorption of the obtained packing material 7 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 8.606 mL, 9.769 mL, 9.9567 mL, 10.703 mL, 11.470 mL, and 12.112 mL, and it was confirmed that there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that no non-specific adsorption was induced.
Example 2
A porous particle (carrier al) was obtained in the same manner as in Example 1, and then a packing material 2 was obtained as follows.
98 g of the carrier α1 was weighed onto a glass filter and thoroughly cleaned with diethylene glycol dimethyl ether.
After cleaning, the porous particle was placed in a 1 L separable flask, 150 g of diethylene glycol dimethyl ether and 150 g (580 mol % based on the glycidyl group) of 1,4-cyclohexanedimethanol were placed in the separable flask, and stirring and dispersion were carried out.
After that, 1.5 ml of a boron trifluoride diethyl ether complex was added, the temperature was raised to 80° C. while stirring at 200 rpm, and the resulting mixture was subjected to the reaction for 4 hours.
The mixture was cooled, then the resulting porous particle (carrier $2) bonded to a diol compound including an alkylene group in the structure thereof was collected by filtration and then washed with 1 L of ion exchanged water to obtain 165 g of a carrier 32.
The progress of the reaction was confirmed by the following procedure.
A part of the dry porous particle into which an alkylene group had been introduced was mixed with potassium bromide, and the resulting mixture was pelletized by applying a pressure and then measured using FT-IR (trade name: Nicolet (registered trademark) iS10, manufactured by Thermo Fisher Scientific Inc.) to check the height of a absorbance peak at 908 cm−1 due to the glycidyl group in the infrared absorption spectrum.
As a result, no absorbance peak at 908 cm−1 was observed by FT-IR.
<Step (C): Introduction Reaction of Glycidyl Group>
150 g of the carrier $2 was weighed onto a glass filter and thoroughly cleaned with dimethylsulfoxide. After cleaning, the carrier $2 was placed in a separable flask, 262.5 g of dimethyl sulfoxide and 150 g of epichlorohydrin were added, the resulting mixture was stirred at room temperature, 37.5 ml of a 30% sodium hydroxide aqueous solution (manufactured by KANTO CHEMICAL CO., INC.) was further added, and the resulting mixture was heated to 30° C. and stirred for 6 hours. After completion of the reaction, the porous particle was transferred onto a glass filter and thoroughly washed with water, acetone, and water in the order presented to obtain 180 g of a porous particle into which a glycidyl group had been introduced (carrier γ2).
The introduction density of the glycidyl group in the obtained carrier γ2 was measured in the same manner as in Example 1. As a result, the density of the glycidyl group was 900 μmol/g.
<Step (D): Introduction Reaction of Polyol>
150 g of the carrier γ2 was weighed onto a glass filter and thoroughly cleaned with diethylene glycol dimethyl ether. After cleaning, the carrier γ2 was placed in a 1 L separable flask, 150 g of diethylene glycol dimethyl ether and 150 g (5760 mol % based on the glycidyl group) of ethylene glycol (log P=−1.36) were placed in the separable flask, and stirring and dispersion were carried out. After that, 1.5 mL of a boron trifluoride diethyl ether complex was added, the temperature was raised to 80° C. while stirring at 200 rpm, and the resulting mixture was subjected to the reaction for 4 hours. The mixture was cooled, and then the reaction product was collected by filtration and washed thoroughly with water to obtain 152 g of a polyol-introduced porous particle (carrier δ2). The carrier δ2 was classified into 16 to 37 μm using a sieve to obtain 140.5 g of a packing material 2.
The alkali resistance of the obtained packing material 2 was evaluated in the same manner as in Example 1. As a result, the amount of a carboxy group produced was 15.2 μmol/mL, and it was confirmed that the packing material 2 had excellent alkali resistance.
Further, the non-specific adsorption of the obtained packing material 2 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 8.814 mL, 9.635 mL, 9.778 mL, 10.37 mL, 10.898 mL, and 12.347 mL, and it was confirmed that there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that no non-specific adsorption was induced.
A packing material 8 was obtained in the same manner as in Example 1 except that 150 g of ethylene glycol was used instead of 1,4-butanediol as an alkylene group-introducing agent.
The alkali resistance of the obtained packing material 8 was evaluated in the same manner as in Example 1. As a result, the amount of a carboxy group produced in the packing material 8 was 108.4 μmol/mL, resulting in poor alkali resistance.
Further, the non-specific adsorption of the obtained packing material 8 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 9.708 mL, 9.8946 mL, 10.6452 mL, 11.5374 mL, and 12.1656 mL, and it was confirmed that there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that no non-specific adsorption was induced.
Example 3
A carrier γ2 was obtained in the same manner as in Example 2.
150 g of the obtained carrier γ2 was weighed onto a glass filter and thoroughly cleaned with diethylene glycol dimethyl ether.
After cleaning, the porous particle was placed in a 1 L separable flask, 150 g of diethylene glycol dimethyl ether and 150 g of polyethylene glycol #200 (manufactured by KANTO CHEMICAL CO., INC., average molecular weight of 190 to 210, log P is unclear, but the close compound tetraethylene glycol (Mw of 194) has a log P of −2.02) (1790 mol % based on glycidyl group) were placed in the separable flask, and stirring and dispersion were carried out.
After that, 1.5 mL of a boron trifluoride diethyl ether complex was added, the temperature was raised to 80° C. while stirring at 200 rpm, and the resulting mixture was subjected to the reaction for 4 hours.
The mixture was cooled, and then the reaction product was collected by filtration and washed thoroughly with water to obtain 152 g of a porous particle into which a polyol had been introduced (carrier 63).
The carrier δ3 was classified into 16 to 37 μm using a sieve to obtain 140.5 g of a packing material 3.
The alkali resistance of the obtained packing material 3 was evaluated in the same manner as in Example 1. As a result, the amount of a carboxy group produced was 16.1 μmol/mL, and it was confirmed that the packing material 3 had excellent alkali resistance.
Further, the non-specific adsorption of the obtained packing material 3 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 8.517 mL, 9.241 mL, 9.47 mL, 10.034 mL, 10.484 mL, and 11.927 mL, and it was confirmed that there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that no non-specific adsorption was induced.
A packing material 9 was obtained in the same manner as in Example 2 except that no glycidyl group was introduced and no polyol was introduced. That is, the carrier $2 obtained in the step (B) of Example 2 was used as the packing material 9.
The non-specific adsorption of the obtained packing material 9 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 8.590 mL, 10.316 mL, 9.603 mL, 10.484 mL, 13.863 mL, and 12.861 mL, and it was confirmed that there was a contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that non-specific adsorption was induced. Because of this, the alkali resistance was not evaluated.
Example 4
A packing material 4 was obtained in the same manner as in Example 3 except that 33.2 g of glycidyl methacrylate (trade name: Blemmer G (registered trademark) manufactured by NOF Corporation), 5.9 g of glycerin-1,3-dimethacrylate (trade name: NK Ester 701, SHIN-NAKAMURA CHEMICAL Co., Ltd.), 58.7 g of diethyl succinate, and 1.9 g of 2,2′-azobis(2,4-dimethylvaleronitrile) were used to provide an oil phase. The amount of glycidyl methacrylate used was 90.0 mol % based on the total amount of the monomers, and the amount of glycerin-1,3-dimethacrylate used was 10.0 mol % based on the total amount of the monomers.
The alkali resistance of the obtained packing material 4 was evaluated in the same manner as in Example 1. As a result, the amount of a carboxy group produced was 11.5 μmol/mL, and it was confirmed that the packing material 4 had excellent alkali resistance.
Further, the non-specific adsorption of the obtained packing material 4 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 7.52 mL, 8.214 mL, 8.451 mL, 9.062 mL, 9.511 mL, and 11.915 mL, and it was confirmed that there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that no non-specific adsorption was induced.
A packing material 10 was obtained in the same manner as in Example 1 except that 150 g (480 mol % based on glycidyl methacrylate) of 1,10-decanediol was used instead of 1,4-butanediol as an alkylene group-introducing agent.
The non-specific adsorption of the obtained packing material 10 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 9.991 mL, 10.15 mL, 10.063 mL, 10.691 mL, 12.172 mL, and 11.531 mL, and it was confirmed that there was a contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that non-specific adsorption was induced. Because of this, the alkali resistance was not evaluated.
Example 5
A packing material 5 was obtained in the same manner as in Example 3 except that 21.5 g of glycidyl methacrylate (trade name: Blemmer G (registered trademark) manufactured by NOF Corporation), 17.6 g of glycerin-1,3-dimethacrylate (trade name: NK Ester 701, SHIN-NAKAMURA CHEMICAL Co., Ltd.), 58.7 g of diethyl succinate, and 1.9 g of 2,2′-azobis(2,4-dimethylvaleronitrile) were used to provide an oil phase.
The amount of glycidyl methacrylate used was 66.2 mol % based on the total amount of the monomers, and the amount of glycerin-1,3-dimethacrylate used was 33.8 mol % based on the total amount of the monomers.
The alkali resistance of the obtained packing material 5 was evaluated in the same manner as in Example 1. As a result, the amount of a carboxy group produced was 18.3 μmol/mL, and it was confirmed that the packing material 5 had excellent alkali resistance.
Further, the non-specific adsorption of the obtained packing material 5 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 8.692 mL, 9.434 mL, 9.625 mL, 10.236 mL, 10.759 mL, and 12.457 mL, and it was confirmed that there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that no non-specific adsorption was induced.
A packing material 11 was obtained in the same manner as in Example 3 except that 13.7 g of glycidyl methacrylate (trade name: Blemmer G (registered trademark) manufactured by NOF Corporation), 25.4 g of glycerin-1,3-dimethacrylate (trade name: NK Ester 701, SHIN-NAKAMURA CHEMICAL Co., Ltd.), 58.7 g of diethyl succinate, and 1.9 g of 2,2′-azobis(2,4-dimethylvaleronitrile) were used to provide an oil phase. The amount of glycidyl methacrylate used was 46.4 mol % based on the total amount of the monomers, and the amount of glycerin-1,3-dimethacrylate used was 53.6 mol % based on the total amount of the monomers.
The non-specific adsorption of the obtained packing material 11 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 8.872 mL, 10.131 mL, 9.82 mL, 10.422 mL, 12.782 mL, and 12.553 mL, and it was confirmed that there was a contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that non-specific adsorption was induced. Because of this, the alkali resistance was not evaluated.
It was confirmed that the exclusion limit molecular weights of the packing materials obtained in Examples 1 to 6 and Comparative Examples 1 to 5 were all 1,000,000 or more.
Example 6
A packing material 6 was obtained in the same manner as in Example 3 except that 33.2 g of glycidyl methacrylate (trade name: Blemmer G (registered trademark) manufactured by NOF Corporation), 5.9 g of ethylene glycol dimethacrylate (trade name: NK Ester 1G, SHIN-NAKAMURA CHEMICAL Co., Ltd.), 29.3 g of butyl acetate, 29.3 g of chlorobenzene, and 1.9 g of 2,2′-azobis(2,4-dimethylvaleronitrile) were used to provide an oil phase. The amount of glycidyl methacrylate used was 88.7 mol % based on the total amount of the monomers, and the amount of ethylene glycol dimethacrylate used was 11.3 mol % based on the total amount of the monomers.
The alkali resistance of the obtained packing material 6 was evaluated in the same manner as in Example 1. As a result, the amount of a carboxy group produced was 12.5 μmol/mL, and it was confirmed that the packing material 6 had excellent alkali resistance.
Further, the non-specific adsorption of the obtained packing material 6 was evaluated in the same manner as in Example 1. As a result, the elution volumes of the samples were 9.613 mL, 10.427 mL, 10.444 mL, 11.066 mL, 11.582 mL, and 12.575 mL, and it was confirmed that there was no contradiction between the order of the molecular weights of the samples and the order of the elution volumes thereof and that no non-specific adsorption was induced.
A packing material 12 was obtained in the same manner as in Example 3 except that 37.1 g of glycidyl methacrylate (trade name: Blemmer G (registered trademark) manufactured by NOF Corporation), 2.0 g of glycerin-1,3-dimethacrylate (trade name: NK Ester 701, SHIN-NAKAMURA CHEMICAL Co., Ltd.), 58.7 g of diethyl succinate, and 1.9 g of 2,2′-azobis(2,4-dimethylvaleronitrile) were used to provide an oil phase. The amount of glycidyl methacrylate used was 96.7 mol % based on the total amount of the monomers, and the amount of glycerin-1,3-dimethacrylate used was 3.3 mol % based on the total amount of the monomers.
Packing into a stainless steel column using the obtained packing material 12 was attempted. However, the back pressure was high, making liquid feeding difficult, and this made it impossible to carry out the packing. Because of this, neither of the evaluations was able to be carried out.
Results of the above Examples and Comparative Examples are shown in Table 1.
From the above results, by adopting the configuration of the present invention, a packing material having suppressed non-specific adsorption and high alkali resistance can be obtained.
When no hydrophobic portion is provided or when the alkylene chain is short, the alkali resistance is low as shown in Comparative Examples 1 and 2. In addition, it was found that when the alkylene chain is too long or when no hydrophilic portion is provided, the hydrophobicity is strong, and non-specific adsorption is induced as shown in Comparative Examples 3 and 4. In addition, in Comparative Example 5 having many repeating units derived from a polyfunctional monomer, it was found that non-specific adsorption was induced, and in Comparative Example 6 having fewer repeating units derived from a polyfunctional monomer, it was found that the back pressure applied to the apparatus was high, making column packing difficult.
Full text: Click here
A 300
Acetone
Adsorption
Alkalies
Anabolism
Aprotinin
boron trifluoride
Buffers
butyl acetate
butylene
Butylene Glycols
chlorobenzene
COMP protocol
Cyclohexane
cyclohexanedimethanol
diethylamine
diethyl succinate
diglyme
Epichlorohydrin
Esters
Ethanol
ethylene dimethacrylate
Ethylenes
Ethyl Ether
Filtration
G 130
gamma-Globulin
Gel Chromatography
Glycerin
glycidyl methacrylate
Glycol, Ethylene
High-Performance Liquid Chromatographies
Hydrochloric acid
Hydrolysis
Nitrogen
Polyethylene Glycols
Polymers
polyol
Polyvinyl Alcohol
potassium bromide
Potassium Chloride
potassium hydroxide
Pressure
Ribonucleases
Sodium Hydroxide
sodium phosphate
Solvents
Sorbitol
Stainless Steel
Sulfoxide, Dimethyl
tetraethylene glycol
Thyroglobulin
Titrimetry
Uridine
Example 2
A sample volume of creatinine sample of 300 uL is placed on the electrode having the FeCl3 receptor of 0.6 mg then the peak reduction current value is noted from cyclic voltammogram specifying a potential window from 0.6 V to −1.0 V with scan rate of 0.1 V/sec in CHI Electrochemical workstation. The value of peak reduction current is measured as 105 μA. The presence of this current value is searched in the values as provided in Table 1 and the corresponding concentration of urine creatinine is retrieved, which is 240 mg/dL.
Full text: Click here
A 300
Creatinine
Radionuclide Imaging
Urine
Example 4
A sample volume of creatinine sample of 300 uL is placed on the electrode having the MB-FeCl3 receptor of 0.6 mg and then the peak reduction current value is observed from cyclic voltammogram by varying a potential window from 0.6 V to −1.0 V, with scan rate of 0.1 V/sec in CHI-Electrochemical workstation. The value of peak reduction current is noted 110 μA. The presence of this current value is searched in the Table 2 and the corresponding concentration of urine creatinine is obtained is 373 mg/dL.
Full text: Click here
A 300
Creatinine
Radionuclide Imaging
Urine
Example 5
Both end dimethylvinylsiloxy-capped dimethylpolysiloxane having an average DOP of 1,800, 65 parts, was mixed with 40 parts of fumed silica having a BET specific surface area of 300 m2/g (Aerosil 300 by Nippon Aerosil Co.), 8 parts of hexamethyldisilazane, 0.1 part of 1,3-divinyl-1,1,3,3-tetramethyldisilazane (vinyl content 0.0116 mol/g), and 2.0 parts of water at 25° C. for 30 minutes. The mixture was heated at 150° C., continuously stirred for 3 hours, and cooled, obtaining a silicone rubber base. This silicone rubber base had a very high viscosity and was difficult to handle, with any further study interrupted.
Full text: Click here
1,3-butadiene
A 300
Aerosil
dimethicone
Feelings
Fingers
hexamethyldisilazane
n-hexane
Polyvinyl Chloride
Silicon Dioxide
Silicone Elastomers
Tears
Touch
Viscosity
Top products related to «A 300»
Sourced in Germany, United States, United Kingdom, Netherlands, Spain, Japan, Canada, France, China, Australia, Italy, Switzerland, Sweden, Belgium, Denmark, India, Jamaica, Singapore, Poland, Lithuania, Brazil, New Zealand, Austria, Hong Kong, Portugal, Romania, Cameroon, Norway
The RNeasy Mini Kit is a laboratory equipment designed for the purification of total RNA from a variety of sample types, including animal cells, tissues, and other biological materials. The kit utilizes a silica-based membrane technology to selectively bind and isolate RNA molecules, allowing for efficient extraction and recovery of high-quality RNA.
Sourced in United States, China, Japan, Germany, United Kingdom, Canada, France, Italy, Australia, Spain, Switzerland, Netherlands, Belgium, Lithuania, Denmark, Singapore, New Zealand, India, Brazil, Argentina, Sweden, Norway, Austria, Poland, Finland, Israel, Hong Kong, Cameroon, Sao Tome and Principe, Macao, Taiwan, Province of China, Thailand
TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
Sourced in United States, Germany, Canada, China, France, United Kingdom, Japan, Netherlands, Italy, Spain, Australia, Belgium, Denmark, Switzerland, Singapore, Sweden, Ireland, Lithuania, Austria, Poland, Morocco, Hong Kong, India
The Agilent 2100 Bioanalyzer is a lab instrument that provides automated analysis of DNA, RNA, and protein samples. It uses microfluidic technology to separate and detect these biomolecules with high sensitivity and resolution.
Sourced in United States, China, Germany, United Kingdom, Spain, Australia, Italy, Canada, Switzerland, France, Cameroon, India, Japan, Belgium, Ireland, Israel, Norway, Finland, Netherlands, Sweden, Singapore, Portugal, Poland, Czechia, Hong Kong, Brazil
The MiSeq platform is a benchtop sequencing system designed for targeted, amplicon-based sequencing applications. The system uses Illumina's proprietary sequencing-by-synthesis technology to generate sequencing data. The MiSeq platform is capable of generating up to 15 gigabases of sequencing data per run.
Sourced in United States, China, Germany, United Kingdom, Hong Kong, Canada, Switzerland, Australia, France, Japan, Italy, Sweden, Denmark, Cameroon, Spain, India, Netherlands, Belgium, Norway, Singapore, Brazil
The HiSeq 2000 is a high-throughput DNA sequencing system designed by Illumina. It utilizes sequencing-by-synthesis technology to generate large volumes of sequence data. The HiSeq 2000 is capable of producing up to 600 gigabases of sequence data per run.
Sourced in United States, China, Germany, United Kingdom, Canada, Switzerland, Sweden, Japan, Australia, France, India, Hong Kong, Spain, Cameroon, Austria, Denmark, Italy, Singapore, Brazil, Finland, Norway, Netherlands, Belgium, Israel
The HiSeq 2500 is a high-throughput DNA sequencing system designed for a wide range of applications, including whole-genome sequencing, targeted sequencing, and transcriptome analysis. The system utilizes Illumina's proprietary sequencing-by-synthesis technology to generate high-quality sequencing data with speed and accuracy.
Sourced in United States, China, United Kingdom, Hong Kong, France, Canada, Germany, Switzerland, India, Norway, Japan, Sweden, Cameroon, Italy
The HiSeq 4000 is a high-throughput sequencing system designed for generating large volumes of DNA sequence data. It utilizes Illumina's proven sequencing-by-synthesis technology to produce accurate and reliable results. The HiSeq 4000 has the capability to generate up to 1.5 terabytes of data per run, making it suitable for a wide range of applications, including whole-genome sequencing, targeted sequencing, and transcriptome analysis.
Sourced in United States, Germany, United Kingdom, Japan, China, France, Denmark, Italy, Spain
The Q Exactive is a high-resolution mass spectrometer designed for accurate and sensitive analysis of a wide range of samples. It features a quadrupole mass filter and an Orbitrap mass analyzer, providing high-resolution, accurate mass measurements for qualitative and quantitative applications.
Sourced in United States, United Kingdom, Germany, China, Canada, Japan, Italy, France, Belgium, Australia, Uruguay, Switzerland, Israel, India, Spain, Denmark, Morocco, Austria, Brazil, Ireland, Netherlands, Montenegro, Poland
Matrigel is a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, a tumor rich in extracellular matrix proteins. It is widely used as a substrate for the in vitro cultivation of cells, particularly those that require a more physiologically relevant microenvironment for growth and differentiation.
Sourced in United States, Germany, China, United Kingdom, Japan, Denmark, Uruguay
The Q Exactive mass spectrometer is a high-resolution, accurate-mass (HRAM) instrument designed for advanced proteomics, metabolomics, and small molecule applications. It combines a quadrupole mass filter with a high-field Orbitrap mass analyzer to provide precise mass measurements and high-quality data.
More about "A 300"
A 300 is a comprehensive, AI-powered platform that assists researchers in optimizing their research protocols for reproducibility and accuracy.
It enables users to easily locate, compare, and identify the best protocols and products for their research projects.
The platform leverages intelligent protocol comparisons from literature, preprints, and patents, providing researchers with streamlined, data-driven decision-making capabilities.
The A 300 platform is designed to simplify the research workflow and ensure robust, reliable results.
It integrates seamlessly with various scientific tools and technologies, such as the RNeasy Mini Kit for RNA extraction, the TRIzol reagent for RNA isolation, the Agilent 2100 Bioanalyzer for sample quality assessment, and the MiSeq, HiSeq 2000, HiSeq 2500, HiSeq 4000, and Q Exactive platforms for next-generation sequencing and mass spectrometry analysis.
With the A 300, researchers can experience a streamlined, data-driven approach to their 300 research projects.
The platform's intelligent protocol comparisons help identify the most suitable methods and products, ensuring reproducibility and accuracy in their experimental designs.
The integration with tools like Matrigex for cell culture and the Q Exactive mass spectrometer for proteomic analysis further enhances the research capabilities of the A 300 platform.
By leveraging the power of AI and data-driven insights, the A 300 empowers researchers to make informed decisions, optimize their workflows, and achieve reliable, high-quality results.
Whether you're working on a single project or managing a portfolio of 300 research initiatives, the A 300 platform is designed to simplify your research journey and drive your scientific discoveries forward.
It enables users to easily locate, compare, and identify the best protocols and products for their research projects.
The platform leverages intelligent protocol comparisons from literature, preprints, and patents, providing researchers with streamlined, data-driven decision-making capabilities.
The A 300 platform is designed to simplify the research workflow and ensure robust, reliable results.
It integrates seamlessly with various scientific tools and technologies, such as the RNeasy Mini Kit for RNA extraction, the TRIzol reagent for RNA isolation, the Agilent 2100 Bioanalyzer for sample quality assessment, and the MiSeq, HiSeq 2000, HiSeq 2500, HiSeq 4000, and Q Exactive platforms for next-generation sequencing and mass spectrometry analysis.
With the A 300, researchers can experience a streamlined, data-driven approach to their 300 research projects.
The platform's intelligent protocol comparisons help identify the most suitable methods and products, ensuring reproducibility and accuracy in their experimental designs.
The integration with tools like Matrigex for cell culture and the Q Exactive mass spectrometer for proteomic analysis further enhances the research capabilities of the A 300 platform.
By leveraging the power of AI and data-driven insights, the A 300 empowers researchers to make informed decisions, optimize their workflows, and achieve reliable, high-quality results.
Whether you're working on a single project or managing a portfolio of 300 research initiatives, the A 300 platform is designed to simplify your research journey and drive your scientific discoveries forward.