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The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes
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Biochemistry
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JoVE Journal Biochemistry
The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes

The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes

DOI:
10.3791/57469-v

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09:47 min

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May 25, 2018

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, ,
1Department of Microbiology and Molecular Cell Biology,Nihon Pharmaceutical University, 2Department of Clinical Pharmaceutics,Nihon Pharmaceutical University

Chapters

  • 00:05Title
  • 00:36Preparation of Tissue Extracts
  • 01:40Digestion of Substrates in Model Protease and Tissue Extracts
  • 02:59Purification of the Iminobiotin-labeled Substrate
  • 05:17Sample Preparation and Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF Mass Spectrometry)
  • 06:48Results: Identification of Protease Specificity
  • 08:46Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol to determine protease specificity in crude mouse tissue extracts using MALDI-TOF spectrometry.

Tags

Protease SpecificityMALDI-TOF Mass SpectrometryPH ManipulationProtease AssayBiotin LabelingStreptavidin Beads

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