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Nanotoxicity pp 133-143 | Cite as

Application of Gelatin Zymography in Nanotoxicity Research

  • Yue Zhang
  • Rong Wan
  • Qunwei Zhang
  • Yiqun MoEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1894)

Abstract

Gelatin zymography is a relatively simple, inexpensive, and powerful technique to detect proteolytic enzymes capable of degrading gelatin from various biological sources. It has been used particularly to detect the two members of the matrix metalloproteinase family, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), due to their potent gelatin-degrading activity. MMP-2 and MMP-9 are also able to degrade a number of extracellular matrix molecules including type IV, V, and XI collagens, laminin, and aggrecan core protein, thus making them important in the nanotoxicity research. In this technique, proteins are separated by sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE), and gelatinases, activated by SDS, digest gelatin embedded in the gel. After staining with Coomassie Brilliant Blue R-250, areas of degradation are visible as clear bands against a blue-stained background. Here, we describe the detailed procedure for gelatin zymography.

Key words

Matrix metalloproteinases MMP-2 MMP-9 Nanoparticles Zymography Gelatin 

Notes

Acknowledgment

This work was partly supported by the NIESH/NIH (ES023693 and ES028911), KSEF-148-502-16-381, Kentucky Lung Cancer Research Program, and an Intramural Research Incentive Grants (50992) from UofL to Dr. Qunwei Zhang.

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Copyright information

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

Authors and Affiliations

  1. 1.Department of Environmental and Occupational Health Sciences, School of Public Health and Information SciencesUniversity of LouisvilleLouisvilleUSA

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