Zymography pp 239-252 | Cite as

Multiplex Cathepsin Zymography to Detect Amounts of Active Cathepsins K, L, S, and V

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1626)

Abstract

Cysteine cathepsins are powerful proteases that can degrade other proteins, among which are the extracellular matrix proteins collagen and elastin. Multiplex cathepsin zymography is an assay that can quantify the amount of active cathepsins in a cell or tissue preparation. This method works for measuring the amounts of active cathepsins K, L, S, and V in a cell or tissue preparation without requiring the use of antibodies for specific identification which tremendously reduces cost. This chapter will demonstrate the utility and interpretation of this method with mammalian cells and tissue to quantify amounts of active cathepsins K, L, S, and V without complicating signals of the procathepsin. Multiplex cathepsin zymography has many advantages: (1) it separates cathepsins K, L, S, and V by electrophoretic migration distance, (2) allows visual confirmation of cathepsin identity, (3) does not detect procathepsins, and (4) can be quantified with densitometry.

Key words

Cathepsins Proteases Cancer Cardiovascular disease Enzymology 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Wallace H. Coulter Department of Biomedical EngineeringGeorgia Institute of Technology and Emory UniversityAtlantaUSA

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