OFFGEL and GELFrEE fractionation: Novel liquid-phase protein recovery strategies in proteomics studies

2021 ◽  
pp. 116282
Author(s):  
Rituparna Banerjee ◽  
Naveena B. Maheswarappa ◽  
Sowmya Dasoju ◽  
Sushma S. Andhe
Keyword(s):  
Liquid Phase ◽  
Protein Recovery ◽  
Recovery Strategies ◽  
Phase Protein

scholarly journals Assembly and Dynamics of Liquid Phase Protein Droplets Comprised of the DEAD-Box RNA Helicase, LAF-1

2015 ◽  
Vol 108 (2) ◽  
pp. 230a-231a
Author(s):  
Shana Elbaum ◽  
Younghoon Kim ◽  
Sua Myong ◽  
Clifford Brangwynne
Keyword(s):  
Liquid Phase ◽  
Rna Helicase ◽  
Dead Box ◽  
Phase Protein ◽  
The Dead

scholarly journals The Utilization of Triton X-100 for Enhanced Two-Dimensional Liquid-Phase Proteomics

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Mina Kim ◽  
Sang-Hee Lee ◽  
Jiho Min ◽  
Fumihisa Kobayashi ◽  
Hyun-Ju Um ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Liquid Phase ◽  
Ribonuclease A ◽  
Protein Profiling ◽  
Protein Recovery ◽  
Recovery Efficiency ◽  
Two Dimensional ◽  
Protein Fractionation ◽  
High Level ◽  
Triton X

One of the main challenges in proteomics lies in obtaining a high level of reproducible fractionation of the protein samples. Automated two-dimensional liquid phase fractionation (PF2D) system manufactured by Beckman Coulter provides a process well suited for proteome studies. However, the protein recovery efficiency of such system is low when a protocol recommended by the manufacturer is used for metaproteome profiling of environmental sample. In search of an alternative method that can overcome existing limitations, this study replaced manufacturer's buffers with Triton X-100 during the PF2D evaluation ofEscherichia coliK12. Three different Triton X-100 concentrations—0.1%, 0.15%, and 0.2%—were used for the first-dimension protein profiling. As the first-dimension result was at its best in the presence of 0.15% Triton X-100, second-dimension protein fractionation was performed using 0.15% Triton X-100 and the standard buffers. When 0.15% Triton X-100 was used, protein recovery increased as much as tenfold. The elution reliability of 0.15% Triton X-100 determined with ribonuclease A, insulin,α-lactalbumin, trypsin inhibitor, and cholecystokinin (CCK) affirmed Triton X-100 at 15% can outperform the standard buffers without having adverse effects on samples. This novel use of 0.15% Triton X-100 for PF2D can lead to greater research possibilities in the field of proteomics.

scholarly journals A rapid solid-phase protein microsequencer

1986 ◽  
Vol 237 (1) ◽  
pp. 73-84 ◽  
Author(s):  
J E Walker ◽  
I M Fearnley ◽  
R A Blows
Keyword(s):  
Liquid Phase ◽  
Solid Phase ◽  
Reaction Chamber ◽  
Covalent Attachment ◽  
Transporter Protein ◽  
Phase Protein ◽  
Reaction Column ◽  
Polar Derivatives ◽  
Protein Sequencer

A solid-phase protein microsequencer is described that has been designed to determine protein sequences with subnanomolar quantities of protein. Its utility has been demonstrated by the determination of many sequences in subunits of mitochondrial F1-ATPase, in a protein isolated from mouse gap junctions and in the mitochondrial phosphate-transporter protein. It has a number of advantages over liquid- and gas-phase sequencers. Firstly, the degradation cycle takes 24 min, more than twice as fast as any other sequencer. This helps to reduce exposure of proteins to inimical reagents and increases throughput of samples. Secondly, polar amino acids such as phosphoserine, and polar derivatives formed by active-site photoaffinity labelling with 8-azido-ATP, are recovered quantitatively from the reaction column and can be positively identified. In other types of sequencer these polar derivatives, being somewhat insoluble in butyl chloride, tend to remain in the reaction chamber of the instrument and so are more difficult to identify. The solid-phase protein sequencer is also more suited than the liquid-phase instrument for analysis of proteolipids from membranes. These hydrophobic proteins tend to dissolve in organic solvents during washing steps in the liquid-phase instrument and are lost. Covalent attachment as used in the solid-phase instrument solves this problem.

Identification and Characterization of Sulfolobus solfataricus P2 Proteome Using Multidimensional Liquid Phase Protein Separations

2008 ◽  
Vol 7 (6) ◽  
pp. 2253-2261 ◽  
Author(s):  
Bobby F. Assiddiq ◽  
Ambrosius P. L. Snijders ◽  
Poh Kuan Chong ◽  
Phillip C. Wright ◽  
Mark. J. Dickman
Keyword(s):  
Liquid Phase ◽  
Sulfolobus Solfataricus ◽  
Protein Separations ◽  
Phase Protein ◽  
Identification And Characterization

scholarly journals Multidimensional liquid phase protein separations in conjunction with stable isotope labelling for quantitative proteomics

PROTEOMICS ◽  
2007 ◽  
Vol 7 (21) ◽  
pp. 3826-3834 ◽  
Author(s):  
Bobby F. Assiddiq ◽  
James C. Williamson ◽  
Ambrosius P. L. Snijders ◽  
Ken Cook ◽  
Mark J. Dickman
Keyword(s):  
Liquid Phase ◽  
Stable Isotope ◽  
Quantitative Proteomics ◽  
Protein Separations ◽  
Stable Isotope Labelling ◽  
Isotope Labelling ◽  
Phase Protein

Elucidation of block boundaries as the dissolution channels in hydrated cement

1978 ◽  
Vol 36 (1) ◽  
pp. 484-485
Author(s):  
N.V. Belov ◽  
U.I. Papiashwili ◽  
B.E. Yudovich
Keyword(s):  
Liquid Phase ◽  
Grain Boundaries ◽  
Benzoyl Peroxide ◽  
Phase Contrast ◽  
Active Role ◽  
Point Of View ◽  
Hardened Cement ◽  
Hydrated Cement ◽  
Hardened Cement Paste

It has been almost universally adopted that dissolution of solids proceeds with development of uniform, continuous frontiers of reaction.However this point of view is doubtful / 1 /. E.g. we have proved the active role of the block (grain) boundaries in the main phases of cement, these boundaries being the areas of hydrate phases' nucleation / 2 /. It has brought to the supposition that the dissolution frontier of cement particles in water is discrete. It seems also probable that the dissolution proceeds through the channels, which serve both for the liquid phase movement and for the drainage of the incongruant solution products. These channels can be appeared along the block boundaries.In order to demonsrate it, we have offered the method of phase-contrast impregnation of the hardened cement paste with the solution of methyl metacrylahe and benzoyl peroxide. The viscosity of this solution is equal to that of water.

Identification of hepatitis a virus in cell cultures by solid phase immune electron microscopy

1986 ◽  
Vol 44 ◽  
pp. 238-239
Author(s):  
C.D. Humphrey ◽  
T.L. Cromeans ◽  
E.H. Cook ◽  
D.W. Bradley
Keyword(s):  
Electron Microscopy ◽  
Liquid Phase ◽  
Cell Cultures ◽  
Rapid Detection ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Immune Electron Microscopy ◽  
Detection And Identification

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.

Microanalysis of silicate glass films grown on α-Al2O3 by pulsed-laser deposition

1993 ◽  
Vol 51 ◽  
pp. 438-439
Author(s):  
Michael P. Mallamaci ◽  
James Bentley ◽  
C. Barry Carter
Keyword(s):  
Liquid Phase ◽  
Single Crystal ◽  
Pulsed Laser Deposition ◽  
Pulsed Laser ◽  
Ceramic Materials ◽  
Laser Deposition ◽  
Triple Junctions ◽  
Ion Milling ◽  
Multicomponent Oxides ◽  
Glass Films

Glass-oxide interfaces play important roles in developing the properties of liquid-phase sintered ceramics and glass-ceramic materials. Deposition of glasses in thin-film form on oxide substrates is a potential way to determine the properties of such interfaces directly. Pulsed-laser deposition (PLD) has been successful in growing stoichiometric thin films of multicomponent oxides. Since traditional glasses are multicomponent oxides, there is the potential for PLD to provide a unique method for growing amorphous coatings on ceramics with precise control of the glass composition. Deposition of an anorthite-based (CaAl2Si2O8) glass on single-crystal α-Al2O3 was chosen as a model system to explore the feasibility of PLD for growing glass layers, since anorthite-based glass films are commonly found in the grain boundaries and triple junctions of liquid-phase sintered α-Al2O3 ceramics.Single-crystal (0001) α-Al2O3 substrates in pre-thinned form were used for film depositions. Prethinned substrates were prepared by polishing the side intended for deposition, then dimpling and polishing the opposite side, and finally ion-milling to perforation.

Effect of furnace atmosphere and silica content on the consolidation behaviour of magnesia partially stabilised zirconia

1990 ◽  
Vol 48 (4) ◽  
pp. 1056-1057
Author(s):  
J. Drennan ◽  
R.H.J. Hannink ◽  
D.R. Clarke ◽  
T.M. Shaw
Keyword(s):  
Liquid Phase ◽  
Silica Content ◽  
Liquid Phase Sintering ◽  
Furnace Atmosphere ◽  
Boundary Phase ◽  
Analytical Electron ◽  
Analytical Electron Microscope ◽  
Series Of Experiments ◽  
Compositional Gradients ◽  
Mobile Liquid

Magnesia partially stabilised zirconia (Mg-PSZ) ceramics are renowned for their excellent nechanical properties. These are effected by processing conditions and purity of starting materials. It has been previously shown that small additions of strontia (SrO) have the effect of removing the major contaminant, silica (SiO2).The mechanism by which this occurs is not fully understood but the strontia appears to form a very mobile liquid phase at the grain boundaries. As the sintering reaches the final stages the liquid phase is expelled to the surface of the ceramic. A series of experiments, to examine the behaviour of the liquid grain boundary phase, were designed to produce compositional gradients across the ceramic bodies. To achieve this, changes in both silica content and furnace atmosphere were implemented. Analytical electron microscope techniques were used to monitor the form and composition of the phases developed. This paper describes the results of our investigation and the presentation will discuss the work with reference to liquid phase sintering of ceramics in general.

Microstructure and microanalysis of sintered Fe-Nd-B permanent magnets

1990 ◽  
Vol 48 (4) ◽  
pp. 990-991
Author(s):  
Mahesh Chandramouli
Keyword(s):  
Electron Microscope ◽  
Liquid Phase ◽  
Permanent Magnets ◽  
Phase Distribution ◽  
Energy Dispersive Spectrometer ◽  
Nucleation And Growth ◽  
Liquid Phase Sintering ◽  
Domain Wall Energy ◽  
Oxide Phases ◽  
Scanning Electron

Magnetization reversal in sintered Fe-Nd-B, a complex, multiphase material, occurs by nucleation and growth of reverse domains making the isolation of the ferromagnetic Fe14Nd2B grains by other nonmagnetic phases crucial. The magnets used in this study were slightly rich in Nd (in comparison to Fe14Nd2B) to promote the formation of Nd-oxides at multigrain junctions and incorporated Dy80Al20 as a liquid phase sintering addition. Dy has been shown to increase the domain wall energy thus making nucleation more difficult while Al is thought to improve the wettability of the Nd-oxide phases.Bulk polished samples were examined in a JEOL 35CF scanning electron microscope (SEM) operated at 30keV equipped with a Be window energy dispersive spectrometer (EDS) detector in order to determine the phase distribution.

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