Abstract
Promoter-driven GUS (beta-glucuronidase) activity is the most commonly used technique for tissue-specific expression patterns in Arabidopsis. In this procedure, GUS enzyme converts 5-bromo-4-chloro-3-indolyl glucuronide (X-Gluc) to a blue product. The staining is very sensitive. Processed samples can be examined under dissecting microscope or Differential Interference Contrast (Nomaski) microscope for bright blue color over cleared transparent background. Note this assay does not provide accurate information to subcellular levels.
Keywords: Gene expression, GUS activity, Histostaining, Arabidopsis, Promoter activity
Materials and Reagents
Equipment
Procedure
Recipes
References
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Cold acetone is for fixation, a way to preserve the cell/tissue structures as in vivo.
Ferro- and Ferri-cyanides form a system buffering redox status (mechanistically like pH buffer). The hydrolyzed indolyl half of X-Gluc by GUS needs to be oxidized to radical before it can form the dimerized blue precipitate as so-called GUS staining.