The Experts below are selected from a list of 270 Experts worldwide ranked by ideXlab platform
George Harauz - One of the best experts on this subject based on the ideXlab platform.
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Ribosomal proteins of Thermomyces lanuginosus--characterisation by Two-Dimensional Gel Electrophoresis and differential disassembly.
Molecular and cellular biochemistry, 1995Co-Authors: Daniel R. Beniac, George HarauzAbstract:One- and Two-Dimensional Gel Electrophoresis were employed to characterise the proteins derived from the ribosomes of the thermophilic fungusThermomyces lanuginosus. Approximately 32 (29 basic and 3 acidic) and 45 (43 basic and 2 acidic) protein spots were resolved fromTh. lanuginosus small and large ribosomal subunits, respectively. The molecular weight of the small subunit proteins ranged from 9,800–36,000 Da with a number average molecular weight of 20,300 Da. The molecular weight range for the large subunit proteins was 12,000–48,500 Da with a number average molecular weight of 25,900 Da. Most proteins appeared to be present in unimolar amounts. These data are comparable with but not identical to those from other eukaryotic ribosomes. The sensitivities of the ribosomal proteins to increasing concentrations of NH4Cl were also evaluated by Two-Dimensional Gel Electrophoresis. Most ribosomal proteins were gradually released over a wide range of salt concentrations but some were preferentially enriched in one or two salt conditions.
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Ribosomal proteins ofThermomyces lanuginosus — characterisation by Two-Dimensional Gel Electrophoresis and differential disassembly
Molecular and Cellular Biochemistry, 1995Co-Authors: Daniel R. Beniac, George HarauzAbstract:One- and Two-Dimensional Gel Electrophoresis were employed to characterise the proteins derived from the ribosomes of the thermophilic fungus Thermomyces lanuginosus . Approximately 32 (29 basic and 3 acidic) and 45 (43 basic and 2 acidic) protein spots were resolved from Th. lanuginosus small and large ribosomal subunits, respectively. The molecular weight of the small subunit proteins ranged from 9,800–36,000 Da with a number average molecular weight of 20,300 Da. The molecular weight range for the large subunit proteins was 12,000–48,500 Da with a number average molecular weight of 25,900 Da. Most proteins appeared to be present in unimolar amounts. These data are comparable with but not identical to those from other eukaryotic ribosomes. The sensitivities of the ribosomal proteins to increasing concentrations of NH_4Cl were also evaluated by Two-Dimensional Gel Electrophoresis. Most ribosomal proteins were gradually released over a wide range of salt concentrations but some were preferentially enriched in one or two salt conditions.
Daniel R. Beniac - One of the best experts on this subject based on the ideXlab platform.
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Ribosomal proteins of Thermomyces lanuginosus--characterisation by Two-Dimensional Gel Electrophoresis and differential disassembly.
Molecular and cellular biochemistry, 1995Co-Authors: Daniel R. Beniac, George HarauzAbstract:One- and Two-Dimensional Gel Electrophoresis were employed to characterise the proteins derived from the ribosomes of the thermophilic fungusThermomyces lanuginosus. Approximately 32 (29 basic and 3 acidic) and 45 (43 basic and 2 acidic) protein spots were resolved fromTh. lanuginosus small and large ribosomal subunits, respectively. The molecular weight of the small subunit proteins ranged from 9,800–36,000 Da with a number average molecular weight of 20,300 Da. The molecular weight range for the large subunit proteins was 12,000–48,500 Da with a number average molecular weight of 25,900 Da. Most proteins appeared to be present in unimolar amounts. These data are comparable with but not identical to those from other eukaryotic ribosomes. The sensitivities of the ribosomal proteins to increasing concentrations of NH4Cl were also evaluated by Two-Dimensional Gel Electrophoresis. Most ribosomal proteins were gradually released over a wide range of salt concentrations but some were preferentially enriched in one or two salt conditions.
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Ribosomal proteins ofThermomyces lanuginosus — characterisation by Two-Dimensional Gel Electrophoresis and differential disassembly
Molecular and Cellular Biochemistry, 1995Co-Authors: Daniel R. Beniac, George HarauzAbstract:One- and Two-Dimensional Gel Electrophoresis were employed to characterise the proteins derived from the ribosomes of the thermophilic fungus Thermomyces lanuginosus . Approximately 32 (29 basic and 3 acidic) and 45 (43 basic and 2 acidic) protein spots were resolved from Th. lanuginosus small and large ribosomal subunits, respectively. The molecular weight of the small subunit proteins ranged from 9,800–36,000 Da with a number average molecular weight of 20,300 Da. The molecular weight range for the large subunit proteins was 12,000–48,500 Da with a number average molecular weight of 25,900 Da. Most proteins appeared to be present in unimolar amounts. These data are comparable with but not identical to those from other eukaryotic ribosomes. The sensitivities of the ribosomal proteins to increasing concentrations of NH_4Cl were also evaluated by Two-Dimensional Gel Electrophoresis. Most ribosomal proteins were gradually released over a wide range of salt concentrations but some were preferentially enriched in one or two salt conditions.
Sean R Gallagher - One of the best experts on this subject based on the ideXlab platform.
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Two-Dimensional Gel Electrophoresis.
Current protocols in molecular biology, 2004Co-Authors: Lonnie D Adams, Sean R GallagherAbstract:Two-Dimensional Gel Electrophoresis is the combination of two high-resolution electrophoretic procedures (isoelectric focusing and SDS-polyacrylamide Gel Electrophoresis) to provide much greater resolution than either procedure alone. In the first-dimension Gel, solubilized proteins are separated according to their isoelectric point (pI) by isoelectric focusing. This Gel is then applied to the top of an SDS-slab Gel and electrophoresed. The proteins in the first-dimension Gel migrate into the second-dimension Gel where they are separated on the basis of their molecular weight. The basic protocols in this unit are based on the type of equipment originally described by O'Farrell in 1975. For very basic or very acidic proteins, two alternate protocols are provided. A third alternate protocol describes how Two-Dimensional Electrophoresis can be performed using a miniGel system. Protein sample preparation is presented in the support protocol.
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Two‐Dimensional Gel Electrophoresis Using the O'Farrell System
Current Protocols in Molecular Biology, 2001Co-Authors: Lonnie D Adams, Sean R GallagherAbstract:Two-Dimensional Gel Electrophoresis is the combination of two high-resolution electrophoretic procedures (isoelectric focusing and SDS-polyacrylamide Gel Electrophoresis) to provide much greater resolution than either procedure alone. In the first-dimension Gel, solubilized proteins are separated according to their isoelectric point (pI) by isoelectric focusing. This Gel is then applied to the top of an SDS-slab Gel and electrophoresed. The proteins in the first-dimension Gel migrate into the second-dimension Gel where they are separated on the basis of their molecular weight. The basic protocols in this unit are based on the type of equipment originally described by O'Farrell in 1975. For very basic or very acidic proteins, two alternate protocols are provided. A third alternate protocol describes how Two-Dimensional Electrophoresis can be performed using a miniGel system. Protein sample preparation is presented in the support protocol.
David W. Speicher - One of the best experts on this subject based on the ideXlab platform.
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Current Protocols in Protein Science - Two-Dimensional Gel Electrophoresis.
Current Protocols in Cell Biology, 1999Co-Authors: Sandra L. Harper, J Mozdzanowski, David W. SpeicherAbstract:While one-dimensional SDS-PAGE separates proteins on the basis of size, Two-Dimensional Gel Electrophoresis separates proteins first on the basis of isoelectric point, then on the basis of size. This method is capable of resolving 1000 to 2000 separate proteins when combined with sensitive detection methods. This unit describes methods for characterizing cell lysates by Two-Dimensional Gel Electrophoresis, including modifications for acidic and basic proteins, the use of immobilized pH gradients, and nonreducing/reducing electrophoretic separations. In addition there are support protocols for determining pH profiles of Gels, casting Immobiline Gels, preparing cell and tissue samples for isoelectric focusing, preparing molecular weight standards, and using Two-Dimensional protein databases.
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Two-Dimensional Gel Electrophoresis.
Current Protocols in Protein Science, 1998Co-Authors: Sandra L. Harper, J Mozdzanowski, David W. SpeicherAbstract:Two-Dimensional Gel Electrophoresis combines two different electrophoretic separating techniques in perpendicular directions to provide a much greater separation of complex protein mixtures than either of the individual procedures. Variations of the most common Two-Dimensional technique are described in this unit, namely isoelectrofocusing (IEF) and SDS-PAGE. This unit also includes support protocols describing pI standards and pH profile measurements, casting Immobiline Gels, preparation of tissue culture cells and solid tissues for isoelectricfocusing, preparation of molecular weight standards for Two-Dimensional Gels, and Two-Dimensional protein databases.
Christelle Monribot-espagne - One of the best experts on this subject based on the ideXlab platform.
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Two-Dimensional Gel Electrophoresis of total yeast proteins.
Methods in molecular biology (Clifton N.J.), 2006Co-Authors: Hélian Boucherie, Christelle Monribot-espagneAbstract:Two-Dimensional Gel Electrophoresis (2-DE) offers the opportunity of separating several hundred proteins from a total yeast cellular extract. A detailed description is provided here of the different steps required for the separation and visualization of radiolabeled yeast proteins on high-resolution (24 cm x 20 cm) 2-D Gels. Two methods of protein separation are described. They essentially differ by the way proteins are separated in the first dimension. One is based on the use of isoelectric focusing (IEF) Gels (carrier ampholytes) and the other on the use of ready-made IPG Gels (immobilines). These methods allow separating soluble proteins from a total yeast cellular extract with an isoelectric point ranging between pH 4.0 and 7.0 and a molecular weight ranging between 15,000 and 150,000.
