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X-Gluc

#R0851 0.1g
#R0852 0.5g
Formula: C14H15Br Cl N O7 x C6H13N
Molecular Weight: 521.8

Related Documents (in pdf, ~30-50 KB):

Certificate of Analysis: #R0851, #R0852
MSDS (English)
MSDS (English-USA)
MSDS (German)

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. The glucuronidase cleaves X-Gluc to produce colorless glucuronic acid and an intense blue precipitate of chloro-bromoindigo.

Applications

Quality Control
Greater than 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.

Protocol for the Histochemical Detection of GUS (beta-glucoronidase) (2)

Staining buffer, prepare immediately before use as follows:
Stock solutions Volume per 10 ml staining buffer Final concentration
Water, nuclease-free 830 µl  
1 M sodium phosphate (pH 7.0) 100 µl 0.1 mM
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 ng/ml) in dimethylformamide 20 µl 2 mM

Staining procedure for cells and tissues:

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

References

  1. 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.

  2. Jefferson, R., Assaying chimeric genes in plants: the GUS gene fusion system, plant Mol Biol Rep, 5, 387-405, 1987.
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Updated vasario 27, 2008 12:37