X-Gluc

- Store at -20°C

Catalog#	Size, concentration	Certificate of Analysis	MSDS

R0851	0.1 g	R0851
R0852	0.5 g	R0852

Product information

X-Gluc
Formula: C₁₄H₁₃BrClNO₇ x C₆N₁₃N
Molecular weight: 521.8

Applications

Detection of GUS expression in plant cells and tissues (1).
Detection of infections caused by E.coli.
Detection of bacterial contamination in food and water samples.

Description

X-Gluc (5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid, cyclohexylammonium salt) is a substrate for beta-glucuronidase (GUS) which is encoded by gusA, a widely used reporter gene. Glucuronidase cleaves X-Gluc to produce colorless glucuronic acid and an intense blue precipitate of chloro-bromoindigo (see the scheme below).

Quality Control

Purity of ≥ 97% confirmed by HPLC.

Note

Preparation of a 0.1 M stock solution in dimethylformamide or dimethylsulfoxide is recommended.

Storage

Store at -20°C in the dark.

Scheme. Glucuronidase cleaves X-Gluc to produce colorless glucuronic acid and blue precipitate of chloro-bromoindigo.

Patents, Licenses, Trademarks

Protocols & recommendations

RECOMMENDATIONS FOR USE

ADDITIONAL PROTOCOLS

Histochemical Detection of GUS (beta-glucoronidase) (1)

Staining buffer, prepare immediately before use as follows:

Stock solutions	Volume per 1 ml staining buffer	Final concentration
Water, nuclease-free	830 µl
1 M sodium phosphate (pH 7.0)	100 µl	0.1 M
0.5 M EDTA, pH 8.0	20 µl	10 mM
10% Triton X-100	10 µl	0.1% (v/v)
50 mM K₃Fe(CN)₆	20 µl	1 mM
0.1 M X-Gluc (50 mg/ml) in dimethylformamide	20 µl	2 mM

Staining procedure for cells and tissues:

Remove media from the cells or tissue.
Immerse cells/tissue in fresh staining buffer.
Incubate cells/tissue for 12-24 hours at 37°C.
Remove the staining buffer.
Wash with several changes of 50% ethanol (up to 12 hours per wash), until the cells/tissue clears.
Count dark blue cells.

Reference

Jefferson, R., Assaying chimeric genes in plants: the GUS gene fusion system, plant Mol Biol Rep, 5, 387-405, 1987.

References

Sambrook, J., Russell, D.W., Molecular Cloning: A Laboratory Manual, the Third edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 16.42, 2001.
Jefferson, R., Assaying chimeric genes in plants: the GUS gene fusion system, plant Mol Biol Rep, 5, 387-405, 1987.