Journal
/
Chemistry
/
A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl
JoVE Journal
Chemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Chemistry
A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl

A Colorimetric Assay that Specifically Measures Granzyme B Proteolytic Activity: Hydrolysis of Boc-Ala-Ala-Asp-S-Bzl

DOI:
10.3791/52419-v

•

05:20 min

•

November 28, 2014

•

, ,
1Cancer Immunology Program,Peter MacCallum Cancer Centre

Chapters

  • 00:05Title
  • 01:00Sample Preparation
  • 02:07Granzyme B Activity Assay
  • 03:30Results: The Ala-Ala-Asp-S-Benzyl Substrate is Specific for Granzyme B
  • 04:53Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a simple, quantitative colorimetric assay that specifically measures the proteolytic activity of human, mouse or rat Granzyme B (GzmB). This protocol can be easily adapted for determining protease activity of other granule serine proteases by the hydrolysis of other synthetic peptide substrates with an appropriate recognition sequence.

Tags

Keywords: Granzyme BColorimetric AssayProteolytic ActivityBoc-Ala-Ala-Asp-S-BzlCytotoxic T LymphocytesNatural Killer CellsSerine ProteaseApoptosisSubstrate SpecificityRecombinant Proteases

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/52419?article_type=v

Rapid Colorimetric Assays to Qualitatively Distinguish RNA and DNA in Biomolecular Samples
check_url/52419?article_type=v

Steady-state, Pre-steady-state, and Single-turnover Kinetic Measurement for DNA Glycosylase Activity
check_url/52419?article_type=v

Fluorescence-based Monitoring of PAD4 Activity via a Pro-fluorescence Substrate Analog
check_url/52419?article_type=v

Dynamic Electrochemical Measurement of Chloride Ions
check_url/52419?article_type=v

A Method to Manipulate Surface Tension of a Liquid Metal via Surface Oxidation and Reduction
check_url/52419?article_type=v

Preparation and Reactivity of a Triphosphenium Bromide Salt: A Convenient and Stable Source of Phosphorus(I)
check_url/52419?article_type=v

Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS
check_url/52419?article_type=v

Microfluidic Preparation of Liquid Crystalline Elastomer Actuators
check_url/52419?article_type=v

Synthesis of Esters Via a Greener Steglich Esterification in Acetonitrile
check_url/52419?article_type=v

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

Read Article