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Matrix-assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) Mass Spectrometric Analysis of Intact Proteins Larger than 100 kDa
JoVE Journal
Chemistry
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JoVE Journal Chemistry
Matrix-assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) Mass Spectrometric Analysis of Intact Proteins Larger than 100 kDa

Matrix-assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) Mass Spectrometric Analysis of Intact Proteins Larger than 100 kDa

DOI:
10.3791/50635-v

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07:49 min

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September 09, 2013

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,
1Institute of Structural Biology “J.P. Ebel”, UMR5075, Commissariat à L’Energie Atomique et aux Energies Alternatives (CEA), Centre National de la Recherche Scientifique (CNRS),Université J. Fourier

Chapters

  • 00:05Title
  • 01:09Recrystallization of Matrices in order to Improve Their Purity (Optional)
  • 02:06Cleaning of MALDI Stainless Steel Target
  • 02:49&#945-CHCA Thin Layer Solution: Preparation and Deposition on the MALDI Target
  • 03:18Preparation of Matrix Solutions: &#945-CHCA, DHB, and CHCA_DHB Mixture
  • 04:04Sample Deposition on the MALDI Target and MALDI Spectra Acquisition
  • 05:17Results: MALDI-TOF Analysis of Proteins Larger than 100 kDa
  • 07:08Conclusion

Summary

Automatic Translation

Automatic Translation

Accurate mass measurement represents an important step during the investigation of proteins. Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) can be used for such determination and its key advantage is the tolerance to salts, detergents and contaminants. Here we illustrate an accessible approach for the analysis of proteins larger than 100 kDa by MALDI-MS.

Tags

MALDI-TOFMass SpectrometryIntact ProteinsProtein AnalysisProtein CharacterizationLarge ProteinsProtein Mass DeterminationSample PreparationMatrixCalibrationLaser EnergyAcquisition Time

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