5-Bromo-4-chloro-3-indolyl-N-acetyl β-D-glucosaminide, 98%

Product Introduction

5-Bromo-4-chloro-3-indolyl-N-acetyl β-D-glucosaminide is a chromogenic substrate commonly used for detecting N-acetyl-β-D-glucosaminidase (NAGase) activity. Upon enzymatic hydrolysis, it releases a halogenated indoxyl intermediate, which is subsequently oxidized to form an insoluble blue precipitate, enabling clear visual detection.

This compound is widely applied in microbiological diagnostics, enzyme activity assays, and histochemical staining, particularly for identifying microorganisms or cells expressing N-acetyl-β-D-glucosaminidase activity.

Due to its high sensitivity and distinct color development, it is an ideal reagent for bacterial detection, colony differentiation, and biochemical assays.

Mechanism of Reaction

5-Bromo-4-chloro-3-indolyl-N-acetyl β-D-glucosaminide functions as a chromogenic enzyme substrate.In enzymatic reactions:

• It is hydrolyzed by N-acetyl-β-D-glucosaminidase (NAGase)
• The glycosidic bond is cleaved, releasing 5-bromo-4-chloro-3-indoxyl
• The indoxyl intermediate is oxidized (typically by oxygen)
• A blue insoluble indigo dye is formed

Key features:

• Color development indicates enzyme activity
• Insoluble precipitate enables localization in cells or colonies
• Suitable for qualitative and semi-quantitative detection

Key Research Applications

1.Enzyme Activity Assays

This compound is widely used for detecting NAGase activity.Typical applications include:

• N-acetyl-β-D-glucosaminidase assays
• Glycosidase activity detection
• Enzyme screening experiments

2.Microbiological Identification

It is commonly used in microbial diagnostics and differentiation.Applications include:

• Detection of NAGase-positive bacteria
• Chromogenic culture media
• Colony color-based identification

3.Histochemical and Cytochemical Staining

The substrate enables visual localization of enzyme activity in biological samples.Applications include:

• Tissue staining
• Cellular enzyme localization
• Microscopy-based detection

4.Diagnostic and Kit Development

Due to its clear chromogenic response, it is widely used in diagnostic reagent development.Applications include:

• In vitro diagnostic assays
• Enzyme-based detection kits
• Clinical microbiology tools

Recommended Experimental Conditions

Conditions may require optimization depending on organism type or enzyme source.

Advantages in Enzyme Assays

• High specificity for NAGase activity
• Strong and visible color development
• Enables direct visual detection
• Suitable for solid and liquid assay systems
• Ideal for microbiological screening

Research Areas

Researchers working in the following fields may benefit from this compound:

• Microbiology and bacterial identification
• Enzymology and glycosidase activity studies
• Clinical and diagnostic microbiology
• Histochemistry and cytochemical staining
• Biochemical assay development and screening
• Life science research involving enzyme detection and visualization

What is this compound used for?
It is used as a chromogenic substrate for detecting N-acetyl-β-D-glucosaminidase activity.

How does the detection work?
Enzymatic hydrolysis releases an indoxyl derivative that forms a blue precipitate upon oxidation.

Can it be used for bacterial identification?
Yes, it is widely used in chromogenic media for differentiating microorganisms.