X-GlcA CHX salt Monohydrate 

X-GlcA CHX salt Monohydrate (5-Bromo-4-chloro-3-indolyl-β-D-glucuronide cyclohexylammonium salt monohydrate, CAS 114162-64-0) is a water-soluble chromogenic substrate optimized for β-glucuronidase (GUS, EC 3.2.1.31) detection in plant molecular biology, microbiology, and histochemistry, hydrolyzing to generate an insoluble intense blue precipitate (dibromo-dichloro-indigo) that precisely localizes GUS expression in tissues, colonies, or plaques. With formula C14H13BrClNO7·C6H13N·H2O (MW 539.80 g/mol), this white crystalline salt pairs the β-D-glucuronopyranuronic acid aglycone with cyclohexylamine counterion for enhanced solubility (>10 mg/mL H2O) versus sodium salt, enabling uniform incorporation into agarose embedding media (0.5-1 mM) for whole-mount plant staining without diffusion artifacts. Enzymatic cleavage by uidA/GUS liberates the indol-3-ol that auto-oxidizes/dimerizes to the intensely colored, membrane-impermeant indigo dye, supporting >99% specific visualization of transgenic events in tobacco, Arabidopsis, and maize transformation workflows. The monohydrate form ensures batch-to-batch consistency for quantitative histochemical scoring (0-5+ scale), while the CHX counterion stabilizes against spontaneous hydrolysis during prolonged incubations (16-48h, 37°C), distinguishing it from less soluble free acid forms. Widely adopted in >50,000 GUS reporter studies since 1990, it excels in tissue clearing protocols, transient expression assays (particle bombardment/PEG), and dual-reporter systems orthogonal to X-Gal/lacZ blue/white screening.

Appearance

• White crystalline powder.
• Hydrolysis yields water-insoluble deep blue precipitate.

Source

• Koenigs-Knorr glycosylation followed by cyclohexylamine salt formation.
• Commercial molecular biology grade (Duchefa >98%, GoldBio).

Molecular Weight and Structure

• Formula: C20H26BrClN2O8; MW 539.80 g/mol (monohydrate).
• Cyclohexylammonium (2R,3S,4S,5S,6S)-6-[(5-bromo-4-chloro-1H-indol-3-yl)oxy]-3,4,5-trihydroxyoxane-2-carboxylate monohydrate.​

Sugar Specificity

• Highly selective β-D-glucuronopyranoside for GUS/β-glucuronidases.
• No cross-reactivity with glucosidases/galactosidases.​

Biological Activity

• Chromogenic GUS reporter; non-toxic to plant cells/bacteria.
• 1000-fold localized color amplification.​

Purity and Microbial Contamination

• ≥98% (HPLC); molecular biology certified.
• Synthetic; non-sterile.​

Identity and Quality Control

• ¹H-NMR verified; water solubility >10 mg/mL.
• Characteristic blue product confirmed post-GUS incubation.​

Shelf Life and Storage

• 3 years at -20°C desiccated; aqueous stable 6 months 4°C.
• Light/moisture protected.​

Applications

• GUS histochemical staining (plant transgenics); uidA reporter assays.
• Bacterial GUS selection, tissue embedding protocols.​

Key Characteristics

• Superior water solubility vs. Na salt; diffusion-free staining.
• Quantitative histochemistry; CHX stabilization.​

Citation Links

• https://www.hepattack.com/en/product03_cid_212_pid_488.html​
• https://pubchem.ncbi.nlm.nih.gov/compound/X-GlcA-CHA-Salt​
• https://duchefa-biochemie.com/product/details/number/X1405​
• https://www.chemicalbook.com/ChemicalProductProperty_IN_CB9313226.htm​
• https://www.chemicalbook.com/ChemicalProductProperty_EN_CB9313226.htm​
• https://www.biosynth.com/Files/MSDS/B7/30/MSDS_B-7300_7000_EN.pdf​
• https://www.sigmaaldrich.com/IN/en/search/x-glca?focus=products​
• https://www.goldbio.com/products/x-gluc-chx-salt​
• http://chemsynlab.com/cn/product/114162-64-0.html​
• https://pubchem.ncbi.nlm.nih.gov/compound/X-GlcA-Sodium-Salt