JoVE Journal
Biochemistry
Biochemistry
This content is Open Access.
Chapters
Summary
This protocol describes the use of high-specificity ion-exchange chromatography with multi-angle light scattering for an accurate molar mass determination of proteins, protein complexes, and peptides in a heterogeneous sample. This method is valuable for quality assessment, as well as for the characterization of native oligomers, charge variants, and mixed-protein samples.
Tags
Keywords: Ion Exchange Chromatography IEX Multi-angle Light Scattering MALS Protein Separation Protein Characterization Protein Quality Control Oligomeric State Homogeneity Identity Conformation Structure Post-translation Modifications Anion Exchange AIEX Cation Exchange CIEX BSA Buffer FPLC DLS
PLAYLIST
Differential Scanning Calorimetry (DSC)
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Education • Materials EngineeringDifferential Scanning Calorimetry -
Research • BiochemistryDifferential Scanning Calorimetry — A Method for Assessing the Thermal Stability and Conformation of Protein Antigen -
Education • General ChemistryUsing Differential Scanning Calorimetry to Measure Changes in Enthalpy
Accelerator Mass Spectrometry (AMS)
Single Crystal X-Ray Diffraction (XRD)
NMR Correlation Spectroscopy (COSY)
Gamma-Ray Spectroscopy (GRS)
Neutron Activation Analysis (NAA)
Laser-Induced Breakdown Spectroscopy (LIBS)
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Research • ChemistryDependence of Laser-induced Breakdown Spectroscopy Results on Pulse Energies and Timing Parameters Using Soil Simulants -
Research • EngineeringQuantitative Analysis of Vacuum Induction Melting by Laser-induced Breakdown Spectroscopy -
Research • EngineeringLaser-induced Breakdown Spectroscopy: A New Approach for Nanoparticle's Mapping and Quantification in Organ Tissue -
Research • EngineeringMeasurement and Analysis of Atomic Hydrogen and Diatomic Molecular AlO, C2, CN, and TiO Spectra Following Laser-induced Optical Breakdown
Ion Mobility Mass Spectrometry (IMMS)
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Research • ChemistryAn HS-MRM Assay for the Quantification of Host-cell Proteins in Protein Biopharmaceuticals by Liquid Chromatography Ion Mobility QTOF Mass Spectrometry -
Research • ChemistryAnalyzing the Photo-oxidation of 2-propanol at Indoor Air Level Concentrations Using Field Asymmetric Ion Mobility Spectrometry
Dynamic Light Scattering (DLS)
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Research • ChemistryApplication of Voltage in Dynamic Light Scattering Particle Size Analysis -
Research • ChemistryMeasurement of Particle Size Distribution in Turbid Solutions by Dynamic Light Scattering Microscopy -
Research • BiochemistryGrowing Protein Crystals with Distinct Dimensions Using Automated Crystallization Coupled with In Situ Dynamic Light Scattering
Cyclic Voltammetry (CV)
Electron Spin Resonance (ESR)
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Education • Inorganic ChemistryElectron Paramagnetic Resonance (EPR) Spectroscopy -
Research • BiologyElectron Spin Resonance Micro-imaging of Live Species for Oxygen Mapping -
Research • BiologyDetection of Nitric Oxide and Superoxide Radical Anion by Electron Paramagnetic Resonance Spectroscopy from Cells using Spin Traps
QuECHERS
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Research • ChemistryPhthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor -
Research • BioengineeringFabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples -
Research • ChemistryGas Chromatography-Mass Spectrometry Paired with Total Vaporization Solid-Phase Microextraction as a Forensic Tool -
Research • EnvironmentA Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants
Photoacoustic Spectroscopy (PAS)
Scanning Tunneling Microscope (STM)
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Research • EngineeringAll-electronic Nanosecond-resolved Scanning Tunneling Microscopy: Facilitating the Investigation of Single Dopant Charge Dynamics -
Research • EngineeringVisualizing Uniaxial-strain Manipulation of Antiferromagnetic Domains in Fe1+YTe Using a Spin-polarized Scanning Tunneling Microscope -
Research • ChemistryProbing the Structure and Dynamics of Interfacial Water with Scanning Tunneling Microscopy and Spectroscopy
Secondary Ion Mass Spectrometry (SIMS)
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Research • ChemistryImaging Corrosion at the Metal-Paint Interface Using Time-of-Flight Secondary Ion Mass Spectrometry -
Research • ChemistryIn Situ Characterization of Hydrated Proteins in Water by SALVI and ToF-SIMS -
Research • ChemistryIn Situ SIMS and IR Spectroscopy of Well-defined Surfaces Prepared by Soft Landing of Mass-selected Ions
Capillary Electrophoresis (CE)
Enzyme-linked Immunosorbent Assay (ELISA)
Ion Exchange Chromatography (IEC)
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Education • Analytical ChemistryIon-Exchange Chromatography -
Research • BiochemistryOnline Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline -
Research • BiochemistryIon Exchange Chromatography (IEX) Coupled to Multi-angle Light Scattering (MALS) for Protein Separation and Characterization
Circular Dichroism (CD)
Super Critical Fluid (SCF) Chromatography
Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS)
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Research • EnvironmentChemical Analysis of Water-accommodated Fractions of Crude Oil Spills Using TIMS-FT-ICR MS -
Research • BiologyMetabolic Pathway Confirmation and Discovery Through 13C-labeling of Proteinogenic Amino Acids -
Research • BiologyDithranol as a Matrix for Matrix Assisted Laser Desorption/Ionization Imaging on a Fourier Transform Ion Cyclotron Resonance Mass Spectrometer -
Research • EnvironmentSingle-throughput Complementary High-resolution Analytical Techniques for Characterizing Complex Natural Organic Matter Mixtures
Carbon-14 Analysis (C-14)
X-Ray Photon Spectroscopy (XPS)
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Education • Materials EngineeringX-ray Photoelectron Spectroscopy -
Research • ChemistryElemental-sensitive Detection of the Chemistry in Batteries through Soft X-ray Absorption Spectroscopy and Resonant Inelastic X-ray Scattering -
Research • ChemistryDetermining the Chemical Composition of Corrosion Inhibitor/Metal Interfaces with XPS: Minimizing Post Immersion Oxidation
Auger Electron Spectroscopy (AES)
Alpha Spectroscopy
Thermal Ionization Mass Spectrometry (TIMS)
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Research • ChemistryAnalysis of Complex Molecules and Their Reactions on Surfaces by Means of Cluster-Induced Desorption/Ionization Mass Spectrometry -
Research • ChemistryCombustion Chemistry of Fuels: Quantitative Speciation Data Obtained from an Atmospheric High-temperature Flow Reactor with Coupled Molecular-beam Mass Spectrometer
Glow Discharge Mass Spectrometry (GDMS)
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Research • ChemistryTotal Internal Reflection Absorption Spectroscopy (TIRAS) for the Detection of Solvated Electrons at a Plasma-liquid Interface -
Research • BiochemistryAnalyzing Dynamic Protein Complexes Assembled On and Released From Biolayer Interferometry Biosensor Using Mass Spectrometry and Electron Microscopy
Ultra-High Performance Liquid Chromatography Tandem Mass Spec (UHPLC–MS/MS)
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Research • BiochemistryQuantitative and Qualitative Method for Sphingomyelin by LC-MS Using Two Stable Isotopically Labeled Sphingomyelin Species -
Research • ChemistrySimultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS -
Research • BiochemistryAn Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
Size-Exclusion Chromatography (SEC)
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Education • BiochemistryChromatography-based Biomolecule Purification Methods -
Research • BiochemistryCharacterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS) -
Research • BiochemistryAnalysis of SEC-SAXS data via EFA deconvolution and Scatter -
Research • ChemistryStandards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS
Polarography
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Research • BiologyHigh-resolution Respirometry to Assess Mitochondrial Function in Permeabilized and Intact Cells -
Research • BiologyRespirometric Oxidative Phosphorylation Assessment in Saponin-permeabilized Cardiac Fibers -
Research • BioengineeringCalorespirometry: A Powerful, Noninvasive Approach to Investigate Cellular Energy Metabolism
