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Showing 1 to 16 of 16 for “"affinity tag"”.
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Optimierte Immobilisierung von Redoxproteinen an funktionale Elektrodenoberflächen für die molekulare Bioelektronik
… heart cyt c on gold electrode surfaces using bioaffinity tags at specified positions within the protein were developed. By using an expression system for cyt c in E. coli and genetic modifications, His and/or Cys tags were incorporated into the amino acid sequence at strategically defined …
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Ribosomal Synthesis of N-methylated peptides
… of the peptide, such as improved target affinity, proteolytic stability or membrane permeability. Therefore synthesis of N-methylated peptide libraries is valuable in screening for drug-like peptides suitable for therapeutic uses. Protein synthesis using recombinant elements (PURE) and …
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G-quadruplex recognition and isolation with small molecules
… Based on the scaffold of this family, an affinity probe was developed, which can mediate the isolation of its nucleic acid targets from human cells. This is the first example of the use of a small molecule with an affinity tag that has been used to isolate a nucleic acid target in a …
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Use of the cellulose-binding domain of a cellulase from cellulomonas fimi for affinity purification of fusion preteins
… domain (CBD) from a bacterial cellulase as an affinity tag for purification of heterologous proteins. The CBD of endoglucanase A (CenA) from Cellulomonas fimi is an N-terminal domain comprising amino acids. CenA binds strongly to cellulose, and the CBD retains this function when separated from …
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Protein Semi-Synthesis in Vivo
… first approach a cellular protein is genetically tagged with one half of a split-intein. The complementary half is linked in vitro to the synthetic probe and this fusion is delivered into cells using a transduction peptide. Association of the intein halves in the cytosol triggers protein …
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Synaptic Protein Profiling in the Mammalian Brain
… of a given neurotransmitter receptor fused to an affinity tag. Immunohistochemistry of fixed brain tissue confirmed the correct localization of each synaptic fusion protein to the appropriate cell type and morphological structure. In order to isolate the synaptic proteins of interest, we developed …
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Cell Wall/Surface Proteome of Candida albicans: its Application in Rapid Identification of Yeast Species by Mass Signature and Characterization by in vitro and in vivo Chemical Labelings
… proteins ("whole cell shaving"), biotin-avidin affinity enrichment of biotinylated peptides, liquid chromatography mass spectrometry (LC-MS) based proteomic approach were employed for unambiguous identification of cell wall/cell wall associated proteins and the exposed peptide segments of these …
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Characterization of cellular stress systems using biological mass spectrometry
… residues were derivatized with an isotope-coded affinity tag (ICAT), thus allowing a direct comparison between the reduced and auto-oxidized forms. Quantitation was based on relative ratios between light and heavy isotopically labeled cysteine containing peptides. The application of the …
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Developing assays to study and screen engineered Escherichia coli NADPH-dependent sulfite reductase
… conditions, facilitating large-scale mutagenic library evaluation through a high-throughput growth selection. Herein, I describe my efforts in developing robust assays for studying engineered E. coli sulfite reductases. To enable recombinant expression of homogeneous protein, I modified …
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Expression, Production, and Purification of Novel Therapeutic Proteins
… coli to express the fusion proteins, and affinity chromatography to isolate the final product.</p> <p>Different medium formulations, feeding strategies, and induction methods were tested in order to develop a production strategy lacking yeast extract or other difficult-to-validate …
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Nanopores, megatonnes, and milliseconds : exploring engineered peptides as antimicrobial, carbon-capture,and biocatalytic agents
… image) enabled characterization of the initial stages of the killing of live Escherichia coli cells. The results suggested that the killing process of CM15 is a combination of a time-variable incubation phase and a more rapid execution phase, offering an interesting parallel between …
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Carbohydrate-Interacting Proteins from Two Nostoc (Cyanobacteria) Species
… enzyme assay and a 32P-labeled affinity tag were utilized to follow the glucuronosyltransferase enzyme through successive purification steps. The active enzyme was extracted from Nostoc commune membrane material using Triton X-100, and then purified by anion exchange …
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A novel in vivo protein refolding technique
… Since the refolded proteins contain an affinity tag, we can purify the properly refolded proteins. This in vivo refolded technique takes the advantage of the high yield prokaryotic expression system and the comprehensive protein folding machinery/QC system of mammalian cells to …
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Establishment of enhanced protein purification strategies for recombinant immunotherapeutics
… strategy by exploring Immobilized Metal Affinity Chromatography (IMAC), Ion Exchange Chromatography (IEX), and Size Exchange Chromatography (SEC) and further strategically integrating these purification techniques to improve protein purity. Intrinsic properties of the proteins of …
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Entwicklung eines Enzym-Modul-Systems zur Synthese und in situ Regeneration von dTDP-aktivierten Desoxyzuckern
… was considerably improved by fusion with a His-Tag because a single IMAC step was sufficient for the production of pure enzyme. The preparative synthesis of dTDP-L-rhamnose starting with the inexpensive substrates dTMP and sucrose was noticeably accelerated by the highly purified enzymes RmlC …
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Multidimensional liquid chromatographic separations for proteomics
… differences. Modifications to the isotope coded affinity tag (ICAT) and isotope tags for relative and absolute quantitation (ITRAQ) protocols were developed to allow protein labeling prior to separation, and the related methodologies ExacTag, isotope coded protein label (ICPL) and stable isotope …