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Enhanced in situ detection of beta-glucuronidase activity in murine tissue

Academic Article
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Overview

authors

  • Kopen, G. C.
  • Prockop, D. J.
  • Phinney, Donald

publication date

  • July 1999

journal

  • Journal of Histochemistry & Cytochemistry  Journal

abstract

  • We outline here a protocol for high-resolution in situ localization of beta-glucuronidase in murine tissues processed in glycol methacrylate (GMA). Murine tissues were first stained with 5-bromo-4-chloro-3-indolyl-beta-d-glucuronic acid (x-gluc), followed by histological processing in GMA. Retention of the blue indigo reaction product after overnight incubations in x-gluc allowed high-resolution localization of beta-glucuronidase activity by brightfield microscopy. When illuminated under darkfield, the x-gluc signal was enhanced, permitting detection even in cells with low-level enzyme activity. This technique offers for the first time a more sensitive enzyme histochemical method of detecting beta-glucuronidase activity in animal tissues and also the opportunity to examine expression at high magni-fication.

subject areas

  • Animals
  • Glucuronates
  • Glucuronic Acid
  • Glucuronidase
  • Immunoenzyme Techniques
  • Liver
  • Methacrylates
  • Mice
  • Spleen
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Research

keywords

  • beta-glucuronidase
  • enzyme histochemistry
  • glycol methacrylate
  • mucopolysaccharidosis type VII
  • x-gluc
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Identity

International Standard Serial Number (ISSN)

  • 0022-1554

PubMed ID

  • 10375385
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Additional Document Info

start page

  • 965

end page

  • 968

volume

  • 47

issue

  • 7

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