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Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)
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Biochemistry
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JoVE Journal Biochemistry
Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)

Characterization of Proteins by Size-Exclusion Chromatography Coupled to Multi-Angle Light Scattering (SEC-MALS)

DOI:
10.3791/59615-v

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10:00 min

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June 20, 2019

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, , ,
1Wyatt Technology Corporation, 2Wolfson Centre for Applied Structural Biology, The Alexander Silberman Institute of Life Science,The Hebrew University of Jerusalem, 3Danyel Biotech Ltd.

Chapters

  • 00:04Title
  • 00:57Preparation of Buffers, System, and Sample, and Loading of Sample
  • 03:42Preparation of the MALS Software and the FPLC Software
  • 05:49Analysis of SEC-MALS BSA Data
  • 08:04Results: SEC-MALS Analysis
  • 09:16Conclusion

Summary

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Automatic Translation

This protocol describes the combination of size exclusion chromatography with multi-angle light scattering (SEC-MALS) for absolute characterization of proteins and complexes in solution. SEC-MALS determines the molecular weight and size of pure proteins, native oligomers, heterocomplexes and modified proteins such as glycoproteins.

Tags

Keywords: SEC-MALSSize-exclusion ChromatographyMulti-angle Light ScatteringMolecular WeightMacromoleculesOligomersComplexesGlycoproteinsMembrane ProteinsAbsolute MethodPeptidesPolymersProteinsNucleocapsidsPolysaccharidesVirus-like ParticlesVirusesPhosphate-buffered SalineFlow RateDRI Flow CellPurge ModeSystem CleanlinessRefractive Index

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