A modified gelatin zymography technique incorporating total protein normalization

Julia Raykin, Eric Snider, Sruti Bheri, John Mulvihill, C. Ross Ethier

Research output: Contribution to journalArticlepeer-review

Abstract

Gelatinase zymography is a commonly used laboratory procedure; however, variability in sample loading and concentration reduce the accuracy of quantitative results obtained from this technique. To facilitate normalization of gelatinase activity by loaded protein amount, we developed a protocol using the trihalocompound 2,2,2-trichloroethanol to allow for gelatin zymography and total protein labeling within the same gel. We showed that detected protein levels increased linearly with loading, and describe a loading concentration range over which normalized gelatinase activity was constant. We conclude that in-gel total protein detection is feasible in gelatin zymography and greatly improves comparison of gelatinase activity between samples.

Original languageEnglish
Pages (from-to)8-10
Number of pages3
JournalAnalytical Biochemistry
Volume521
DOIs
Publication statusPublished - 15 Mar 2017
Externally publishedYes

Keywords

  • Gelatinase
  • Loading controls
  • Matrix metalloproteinases
  • Trihalocompounds
  • Zymography

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