X-Gluc Dicyclohexylamine (5-Bromo-4-chloro-3-indolyl-beta-D-glucuronide cyclohexylammonium salt) is a reagent to detect β-glucuronidase and can be used in molecular biology experiments to mark and select the expression of target genes.

Instructions
I. Solution preparation
1. Stock solution: Dissolve X-Gluc Dicyclohexylamine powder in DMSO (dimethylsulfonamide) or acetone, the stock solution concentration is usually 50-100 mg/mL.
2. Working solution: Dilute the stock solution to a final concentration of 1-2 mg/mL using sodium phosphate buffer (pH 7.0), but the optimal concentration may need to be optimized depending on the application.
II. Histochemical staining procedure
1. Sample preparation: Prepare plant tissues, cells or other samples to be stained. For plant tissues, samples may need to be fixed and permeabilized with a fixative such as formalin or glutaraldehyde.
2. Incubation: Incubate the prepared sample with X-Gluc working solution (1-2 mg/mL), usually at 37°C for several hours (usually 4-16 hours). The incubation time can be adjusted according to the sensitivity of the sample and the intensity of the desired staining.
3. Visualization: After the incubation period, wash the sample to remove excess substrate and it can then be observed under an optical microscope. β-Glucuronidase activity will result in the formation of an indigo-colored precipitate, resulting in a blue stain in the tissue.
4. Counterstain (optional): In some cases, a counterstain such as eosin may be used to highlight other features of the tissue or cells.