The Experts below are selected from a list of 291 Experts worldwide ranked by ideXlab platform
Peter Eisner - One of the best experts on this subject based on the ideXlab platform.
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rapeseed Protein concentrates for non food applications prepared from pre pressed and cold pressed press cake via acidic precipitation and ultrafiltration
Industrial Crops and Products, 2019Co-Authors: Andreas Fetzer, Thomas Herfellner, Peter EisnerAbstract:Abstract Rapeseed Protein concentrates (RPCs), prepared from residual press-cakes, show excellent techno-functional properties, such as emulsification and foaming. However, food application is hindered by residual contents of anti-nutritive components. Limited research has been invested in studying RPCs as a potential ingredient in non-food applications. The present study reports RPC preparation from cold-pressed rapeseed meal (CPM) and pre-pressed rapeseed meal (PPM) as a potential ingredient for the chemical industry. Protein Isolation was achieved after extraction at pH 5.7–7.0 via i) ultrafiltration or ii) a combination of acidic precipitation followed by ultrafiltration. Protein Isolation yields obtained from CPM (36.5%–40.6%) were 60%–90% higher compared to PPM (19.4%–26.0%), highlighting the positive effect of low-temperature processing during defatting. Protein contents of RPCs were 75.3%–87.1% and were highest for preparations obtained through precipitation. RPCs obtained through ultrafiltration showed good solubility properties and very high values for emulsifying capacity (688–768 mL/g) as well as foaming activity (1834%–2834%). In contrast, precipitated RPCs showed lower functional values (410–445 mL/g, 888%–938%, respectively). All RPCs had excellent film-forming properties in cast-film experiments. Thus, RPCs are promising ingredients for industrial non-food applications such as adhesives, detergents, paints, varnishes and biodegradable polymers.
Andreas Fetzer - One of the best experts on this subject based on the ideXlab platform.
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rapeseed Protein concentrates for non food applications prepared from pre pressed and cold pressed press cake via acidic precipitation and ultrafiltration
Industrial Crops and Products, 2019Co-Authors: Andreas Fetzer, Thomas Herfellner, Peter EisnerAbstract:Abstract Rapeseed Protein concentrates (RPCs), prepared from residual press-cakes, show excellent techno-functional properties, such as emulsification and foaming. However, food application is hindered by residual contents of anti-nutritive components. Limited research has been invested in studying RPCs as a potential ingredient in non-food applications. The present study reports RPC preparation from cold-pressed rapeseed meal (CPM) and pre-pressed rapeseed meal (PPM) as a potential ingredient for the chemical industry. Protein Isolation was achieved after extraction at pH 5.7–7.0 via i) ultrafiltration or ii) a combination of acidic precipitation followed by ultrafiltration. Protein Isolation yields obtained from CPM (36.5%–40.6%) were 60%–90% higher compared to PPM (19.4%–26.0%), highlighting the positive effect of low-temperature processing during defatting. Protein contents of RPCs were 75.3%–87.1% and were highest for preparations obtained through precipitation. RPCs obtained through ultrafiltration showed good solubility properties and very high values for emulsifying capacity (688–768 mL/g) as well as foaming activity (1834%–2834%). In contrast, precipitated RPCs showed lower functional values (410–445 mL/g, 888%–938%, respectively). All RPCs had excellent film-forming properties in cast-film experiments. Thus, RPCs are promising ingredients for industrial non-food applications such as adhesives, detergents, paints, varnishes and biodegradable polymers.
Robert Tjian - One of the best experts on this subject based on the ideXlab platform.
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dcas9 targeted locus specific Protein Isolation method identifies histone gene regulators
Proceedings of the National Academy of Sciences of the United States of America, 2018Co-Authors: Chiahao Tsui, Carla Inouye, Michaella J Levy, Laurence Florens, Michael P Washburn, Robert TjianAbstract:Eukaryotic gene regulation is a complex process, often coordinated by the action of tens to hundreds of Proteins. Although previous biochemical studies have identified many components of the basal machinery and various ancillary factors involved in gene regulation, numerous gene-specific regulators remain undiscovered. To comprehensively survey the proteome directing gene expression at a specific genomic locus of interest, we developed an in vitro nuclease-deficient Cas9 (dCas9)-targeted chromatin-based purification strategy, called “CLASP” (Cas9 locus-associated proteome), to identify and functionally test associated gene-regulatory factors. Our CLASP method, coupled to mass spectrometry and functional screens, can be efficiently adapted for isolating associated regulatory factors in an unbiased manner targeting multiple genomic loci across different cell types. Here, we applied our method to isolate the Drosophila melanogaster histone cluster in S2 cells to identify several factors including Vig and Vig2, two Proteins that bind and regulate core histone H2A and H3 mRNA via interaction with their 3′ UTRs.
Thomas Braun - One of the best experts on this subject based on the ideXlab platform.
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microfluidic Protein Isolation and sample preparation for high resolution cryo em
Proceedings of the National Academy of Sciences of the United States of America, 2019Co-Authors: Claudio Schmidli, Stefan Albiez, Luca Rima, Ricardo D. Righetto, Inayatulla Mohammed, Paolo Oliva, Lubomír Kováčik, Henning Stahlberg, Thomas BraunAbstract:High-resolution structural information is essential to understand Protein function. Protein-structure determination needs a considerable amount of Protein, which can be challenging to produce, often involving harsh and lengthy procedures. In contrast, the several thousand to a few million Protein particles required for structure determination by cryogenic electron microscopy (cryo-EM) can be provided by miniaturized systems. Here, we present a microfluidic method for the rapid Isolation of a target Protein and its direct preparation for cryo-EM. Less than 1 μL of cell lysate is required as starting material to solve the atomic structure of the untagged, endogenous human 20S proteasome. Our work paves the way for high-throughput structure determination of Proteins from minimal amounts of cell lysate and opens more opportunities for the Isolation of sensitive, endogenous Protein complexes.
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Microfluidic Protein Isolation and sample preparation for high-resolution cryo-EM
2019Co-Authors: Claudio Schmidli, Stefan Albiez, Luca Rima, Ricardo D. Righetto, Inayatulla Mohammed, Paolo Oliva, Lubomír Kováčik, Henning Stahlberg, Thomas BraunAbstract:High-resolution structural information is essential to understand Protein function. Protein-structure determination needs a considerable amount of Protein, which can be challenging to produce, often involving harsh and lengthy procedures. In contrast, the several thousands to a few million Protein particles required for structure-determination by cryogenic electron microscopy (cryo-EM) can be provided by miniaturized systems. Here, we present a microfluidic method for the rapid Isolation of a target Protein and its direct preparation for cryo-EM. Less than one microliter of cell lysate is required as starting material to solve the atomic structure of the untagged, endogenous human 20S proteasome. Our work paves the way for high-throughput structure determination of Proteins from minimal amounts of cell lysate and opens new opportunities for the Isolation of sensitive, endogenous Protein complexes.
J.p. Dean Goldring - One of the best experts on this subject based on the ideXlab platform.
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Measuring Protein Concentration with Absorbance, Lowry, Bradford Coomassie Blue, or the Smith Bicinchoninic Acid Assay Before Electrophoresis.
Methods in molecular biology (Clifton N.J.), 2018Co-Authors: J.p. Dean GoldringAbstract:Measuring the concentration of Proteins is an essential part of enzyme analysis or serves to monitor Protein yields and losses during Protein Isolation procedures. Decisions on the usefulness of any Protein Isolation procedure depend on knowing the concentration of Proteins before and after a procedure. Protein concentration in solution is generally measured with spectrophotometry in the UV range or in the presence of dyes or copper interacting with the Protein. This review describes absorbance at 280 nm, the Lowry, Bradford (Coomassie Blue), and Smith (bicinchoninic acid) assays for measuring Protein and includes suggestions for optimizing each method.
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Concentrating Proteins by Salt, Polyethylene Glycol, Solvent, SDS Precipitation, Three-Phase Partitioning, Dialysis, Centrifugation, Ultrafiltration, Lyophilization, Affinity Chromatography, Immunoprecipitation or Increased Temperature for Protein Is
Methods in molecular biology (Clifton N.J.), 2018Co-Authors: J.p. Dean GoldringAbstract:In Protein Isolation, drug interaction studies, and proteomic or peptidomic procedures, Protein solutions are often concentrated to remove solvents and undesirable molecules, to separate Protein fractions, or to increase Protein concentrations. Proteins can be concentrated by precipitation from solution with ammonium sulfate, polyethylene glycol, organic solvents, trichloroacetic acid, potassium chloride/sodium dodecyl sulfate thermal denaturation, and three-phase partitioning. Solvents can be removed by passage through a semipermeable barrier where Protein solutions are forced against the barrier in a centrifuge tube or with increased pressure, concentrating Proteins in the remaining solution. The semipermeable barrier can be surrounded by a hygroscopic reagent to draw the solvent across the membrane. Proteins can be concentrated by freeze-drying (lyophilization). Unique ligand interactions with Proteins can be used to select for Proteins by affinity purification or immunoprecipitation. All these methods to concentrate Proteins are discussed.
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Methods to Concentrate Proteins for Protein Isolation, Proteomic, and Peptidomic Evaluation
Methods in molecular biology (Clifton N.J.), 2015Co-Authors: J.p. Dean GoldringAbstract:In Protein Isolation, proteomic, or peptidomic procedures, Protein solutions are often concentrated to remove solvents and undesirable molecules, to separate Protein fractions or to increase Protein concentrations. Proteins can be concentrated by precipitation from solution with ammonium sulfate, polyethylene glycol, organic solvent, trichloroacetic acid, potassium chloride/sodium dodecyl sulfate, and three-phase partitioning. Solvents can be removed by passage through a semipermeable barrier where Protein solutions are forced against the barrier in a centrifuge tube or with increased pressure concentrating Protein in the remaining solution. The semipermeable barrier can be surrounded by a hygroscopic reagent to draw the solvent across the membrane. Proteins can be concentrated by freeze-drying (lyophilization). All these methods to concentrate Proteins are discussed.
