SOP describing how to perform Dynamic Light Scattering (DLS) measurements with Gre2p in cuvettes using a Zetasizer Nano DLS reader.
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Created: 27th Apr 2021 at 12:06
Last updated: 6th Jul 2021 at 07:18
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Version History
Version 3 (latest) Created 6th Jul 2021 at 07:18 by Gudrun Gygli
Added BSA in the treatments section.
Version 2 Created 5th May 2021 at 11:54 by Gudrun Gygli
Corrected typos, included DOIs for Chemotion for NDK and HK (if applicable).
Version 1 (earliest) Created 27th Apr 2021 at 12:06 by Gudrun Gygli
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Projects: Simulation Foundries, CML for thermophysical properties of mixtures, Test Project (dummy), Towards Reproducible Enzyme Modeling
Institutions: University of Stuttgart, Karlsruhe Institute of Technology (KIT)
https://orcid.org/0000-0001-9119-1778Expertise: enzyme kinetics, enzymes, Enzymatic reactions, biochemical enzyme characterization, Biochemistry, molecular simulation, molecular modeling, Programming, Bioinformatics, Computational Biology
Tools: Gromacs, Python, Molecular Dynamics, bash, Biochemistry, Bioinformatics, Biochemistry and protein analysis, Enzyme assay, enzyme kinetics, isothermal titration calorimetry, dynamic light scattering, Spectrophotometry
Polyglot European Scientist. I thrive working in interdisciplinary environments combining the study of enzyme reactions and mechanisms with bioinformatics, molecular modelling, automated data analysis and data stewardship.
Work of the Gygli group...
Projects: Test Project (dummy), Towards Reproducible Enzyme Modeling
Web page: Not specified
Investigation 1: An experimental workflow to provide detailed information of the molecular mechanisms of enzymes is described. This workflow will help in the application of enzymes in technical processes by providing crucial parameters needed to plan, model and implement biocatalytic processes more efficiently. These parameters are homogeneity of the enzyme sample (HES), kinetic and thermodynamic parameters of enzyme kinetics and binding of reactants to enzymes. The techniques used to measure ...
Organisms: Not specified
An experimental workflow to provide detailed information of the molecular mechanisms of enzymes is described. This workflow will help in the application of enzymes in technical processes by providing crucial parameters needed to plan, model and implement biocatalytic processes more efficiently. These parameters are homogeneity of the enzyme sample (HES), kinetic and thermodynamic parameters of enzyme kinetics and binding of reactants to enzymes. The techniques used to measure these properties are ...
Submitter: Gudrun Gygli
Studies: DLS measurements (homogeneity of an enzyme sample), ITC binding experiments, ITC kinetic experiments (enzyme activity), Spectrophotometric Activity Measurements
Assays: Analysis of data from ITC experiments (binding), Analysis of data from ITC experiments (kinetics), Binding of HK to Gre2p (ITC-BIND), Binding of NADP+ to Gre2p in HEPES Buffer (ITC-BIND), Binding of NADP+ to Gre2p in KPi Buffer (ITC-BIND), Binding of NADP+ to Gre2p in PBS Buffer (ITC-BIND), Binding of NADPH to Gre2p in HEPES Buffer (ITC-BIND), Binding of NADPH to Gre2p in KPi Buffer (ITC-BIND), Binding of NADPH to Gre2p in PBS Buffer (ITC-BIND), Binding of NADPH to Gre2p in Tween-KPi Buffer (ITC-BIND), Binding of NDK to Gre2p (ITC-BIND), DLS measurements in 2 buffers, DLS measurements in KPi Buffer and in KPi buffer with Tween added, DLS measurements in KPi buffer with BSA added, Kinetic parameters of Gre2p, Kinetics of the reaction of NDK and NADPH with Gre2p (ITC-MIM) in HEPES ..., Kinetics of the reaction of NDK and NADPH with Gre2p (ITC-MIM) in KPi bu..., Kinetics of the reaction of NDK and NADPH with Gre2p (ITC-MIM) in PBS bu..., Kinetics of the reaction of NDK and NADPH with Gre2p (ITC-MIM) in Tween-..., Kinetics of the reaction of NDK and NADPH with Gre2p (ITC-rSIM) in 3 buf..., Selwyn test of Gre2p, Specific activity of Gre2p
Snapshots: Snapshot 1
Measure homogeneity of an enzyme sample (Gre2p) with DLS (dynamic light scattering).
Submitter: Gudrun Gygli
Investigation: Workflow for characterization of enzymes under ...
Assays: DLS measurements in 2 buffers, DLS measurements in KPi Buffer and in KPi buffer with Tween added, DLS measurements in KPi buffer with BSA added
Snapshots: No snapshots
Homogeneity of the Gre2p samples in KPi Buffer before and after different treatments, including different amounts of tween-20 in the buffer (0.01, 0.1, 1.0%).
Submitter: Gudrun Gygli
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Workflow for characterization of enzymes under ...
Organisms: No organisms
SOPs: Python Workflow for Analysis of Specific Activi..., SOP for DLS measurements
Data files: DLS data for Gre2p in KPi Buffer and in KPi buf...
Snapshots: No snapshots
Homogeneity of the Gre2p samples in HEPES and PBS Buffer before and after different treatments.
Submitter: Gudrun Gygli
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Workflow for characterization of enzymes under ...
Organisms: No organisms
SOPs: Python Workflow for Analysis of Specific Activi..., SOP for DLS measurements
Data files: DLS data for Gre2p in HEPES and PBS buffer
Snapshots: No snapshots
Homogeneity of the Gre2p samples in KPi Buffer before and after different treatments, including different amounts of BSA in the buffer (7.5 µM and 75 µM).
Submitter: Felix Ott
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Workflow for characterization of enzymes under ...
Organisms: No organisms
SOPs: Python Workflow for Analysis of Specific Activi..., SOP for DLS measurements
Data files: DLS data for Gre2p in KPi buffer with BSA added
Snapshots: No snapshots