What is the Staining Protocol for GUSPlus?
Standard GUSPlus/PenGUS staining protocol modified from Jefferson RA (1987) Plant Mol. Biol. Rep. 5:387-405) Standard X-GlcA solution 50mM sodium phosphate buffer pH7.0 10mM EDTA pH8.0 0.1% (v/v) Triton X-100 0.5mg/mL X-GlcA Store at 4°C in dark Cover tissue with standard X-glcA solution, vacuum infiltrate for 5 mins. Incubate at 37°C for up to 16h to develop blue colour of product. Non-destructive GUSPlus/PenGUS staining protocol–more recently developed at CAMBIA This method has been tested with GUSPlus and PenGUS (links to the relevant BioForge pages) on rice callus, tobacco shoots and Arabidopsis seedlings and does not prevent continued growth of these tissues. Note that we do not warrant that it may not be detrimental to other tissues or to individual cells. There are a number of modifications/additions to the standard GUS protocol that can be found in the literature, depending on your tissue, and you can experiment with toxicity. Cover tissue with 200 ug/mL X-glcA in 20mM sodium
