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Reverse hyaluronan substrate gel zymography procedure for the detection of hyaluronidase inhibitors

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Abstract

Little is known of the ubiquitous inhibitors of hyaluronidase, molecules that may be important for the deposition of hyaluronan. A reverse hyaluronan-substrate gel procedure is described here that detects such inhibitors, even in crude biological extracts, and is independent of the catalytic mechanism of the target enzyme. Following electrophoresis, hyaluronan-containing gels are incubated in a hyaluronidase solution. Alcian blue-staining bands indicate hyaluronan protected from degradation and the location of hyaluronidase inhibitors. Coordinated use of hyaluronan substrate gel and reverse substrate gel procedures provides estimates of the number and relative molecular sizes of both enzymes and their inhibitors.

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Authors and Affiliations

  1. Lion Corporation, Kanagawa, 256-0811, Japan

    Kazuhiro Mio

  2. Department of Pathology, School of Medicine, University of California, San Francisco, California, 94143-0506, USA

    Robert Stern

Authors
  1. Kazuhiro Mio
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  2. Robert Stern
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Mio, K., Stern, R. Reverse hyaluronan substrate gel zymography procedure for the detection of hyaluronidase inhibitors. Glycoconj J 17, 761–766 (2000). https://doi.org/10.1023/A:1010928523877

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