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Calpain pp 31-38 | Cite as

Casein Zymography for Analysis of Calpain-1 and Calpain-2 Activity

  • Ashim Kumar Biswas
  • Samarth Tandon
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1915)

Abstract

Casein zymography is used to detect calpain activity in cell or tissue lysates. In this technique, lysates are loaded into a polyacrylamide gel containing casein, and the enzymes within the lysates are electrophoretically separated. The casein embedded in the gel acts as a substrate for calpains in the lysates, and its degradation reflects the activity of these enzymes. Calpain degradation of the casein is visualized as distinct bands that are devoid of dye when stained with Coomassie Brilliant Blue G-250. We describe here how calpains can be extracted from muscle tissue and assayed for activity using this technique. This technique is also generally applicable to lysates from other types of cells or tissues.

Key words

Casein zymography Chicken Muscle tissues Substrate zymography Electrophoresis 

Notes

Acknowledgment

This work is supported by DST (Govt. of India) grant SB/FT/LS-283/2012 and the Indian Council of Agricultural Research, New Delhi (Ministry of Agriculture).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Ashim Kumar Biswas
    • 1
  • Samarth Tandon
    • 1
  1. 1.Division of Post-Harvest TechnologyICAR-Central Avian Research InstituteBareillyIndia

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