Y‑GlcA CHX Salt (CAS 216971‑58‑3), systematically 3‑indolyl‑β‑D‑glucuronic acid cyclohexylammonium salt, is a synthetic β‑D‑glucuronidase‑specific chromogenic substrate in which a 3‑indolyl (indoxyl) aglycone is linked via a β‑glycosidic bond to β‑D‑glucuronic acid and presented as the cyclohexylammonium salt. When hydrolyzed by β‑D‑glucuronidase (EC 3.2.1.31, GUS), the indoxyl moiety is released and oxidizes to form an insoluble yellow‑to‑brownish precipitate, enabling clear visual detection of enzyme activity. The CHX salt form is highly soluble in organic solvents such as DMSO or DMF, allowing preparation of concentrated stock solutions that are then diluted into aqueous media for use in bacterial‑colony screening, GUS‑reporter assays, and histochemical staining of β‑glucuronidase activity in plant and animal tissues. The yellow‑brown hue provides good contrast with blue‑colored backgrounds and can be combined with blue‑forming substrates in multiplex‑detection systems. Y‑GlcA CHX Salt is supplied as a high‑purity, research‑use‑only reagent compatible with standard microbiological and molecular‑biology buffers.
Appearance
Off‑white to pale yellow crystalline or lyophilized (freeze‑dried) powder.
Forms a clear, pale‑yellow solution in DMSO, DMF, or methanol; the true chromogenic color (yellow‑to‑brown) appears only after enzymatic hydrolysis and oxidation.
Source / synthesis
Synthetically produced by coupling 3‑indoxyl with β‑D‑glucuronic acid under glycosylation conditions and then converting the free‑acid form into the cyclohexylammonium salt.
Commercially manufactured by specialty enzyme‑substrate suppliers (e.g., Shenzhen GeneSeqTools, Hepattack Bioscience) as a high‑purity GUS‑assay reagent.
Molecular weight and structure
Molecular formula: C20H28N2O7C20H28N2O7.
Molecular weight: 408.45 g/mol.
Structure: 1H‑indol‑3‑yl β‑D‑glucopyranuronate associated with cyclohexylamine (cyclohexylammonium counterion), with a β‑glycosidic linkage between the indole C3‑O and the D‑glucuronic‑acid ring.
Sugar specificity
Specifically hydrolyzed by β‑D‑glucuronidases (GUS, EC 3.2.1.31) and β‑glucuronide‑cleaving enzymes.
Shows minimal cross‑reactivity with other β‑glycosidases under standard assay conditions, providing high selectivity for β‑glucuronidase activity.
Biological activity
Acts as a chromogenic reporter for β‑glucuronidase‑mediated hydrolysis of β‑D‑glucuronosyl linkages in glucuronides and glycosaminoglycan‑derived substrates.
Enzymatic cleavage releases 3‑indoxyl, which oxidizes and dimerizes into a yellow‑to‑brown, water‑insoluble dye, enabling sensitive detection in bacterial colonies, plant‑tissue GUS assays, and histological sections.
Purity and microbial contamination
Typically supplied as a “high‑purity raw material suitable for scientific research” with ≥95–98% purity indicated by vendor data sheets.
Described as non‑sterile, research‑grade powder with low microbial contamination status, suitable for microbiological and biochemical workflows under aseptic handling.
Identity and quality control
Identity inferred from molecular formula, molecular weight, and spectral data consistent with 3‑indolyl‑β‑D‑glucuronic acid cyclohexylammonium salt.
Quality‑control checks include appearance, solubility, and performance in a defined β‑glucuronidase (e.g., E. coli GUS⁺) activity assay on plates or in solution.
Shelf life and storage
Shelf life is generally 12–24 months when stored as a dry powder at –20 °C in a tightly sealed, desiccated container, protected from moisture and light.
Stock solutions in DMSO or DMF should be prepared immediately before use or stored frozen for short periods to minimize degradation and nonspecific oxidation of the indoxyl moiety.
Application
Screening and enumeration of β‑glucuronidase‑positive bacteria (e.g., indicator strains for E. coli) on selective agar media.
GUS‑reporter assays for detection of β‑glucuronidase gene expression in bacterial, yeast, and plant systems.
Histochemical or cytochemical localization of GUS activity in tissue sections and cell preparations, especially where a yellow/brown dye improves contrast over blue substrates.
Key characteristics
Produces a yellow‑to‑brown insoluble dye upon β‑glucuronidase hydrolysis, providing a color‑distinct alternative to blue‑forming GUS substrates.
Defined β‑D‑glucuronide linkage with a cyclohexylammonium counterion, improving solubility in organic stock solvents.
Crystalline or lyophilized powder with a known molecular formula and weight, suitable for quantitative GUS assays and standardized media formulations.
Reviews
There are no reviews yet.