Chinese Name: L-Proline 7-amido-4-methylcoumarin hydrobromide

English Name: L-Proline 7-amido-4-methylcoumarin hydrobromide

Abbreviation: H-Pro-AMC HBr

CAS Number: 115388-93-7

Molecular Formula: C15H17BrN2O3

Molecular Weight: 353.22

Specifications: 100 mg / 500 mg / 1 g (Custom packaging supported)

Form: Lyophilized powder

Product Number: G12060007

Color: Blue fluorescence

Stock: In stock

Chinese Name: (2S)-N-(4-methyl-2-oxo-2H-1-benzopyran-7-yl)-2-pyrrolidinecarboxamide

English Name: L-Proline 7-amido-4-methylcoumarin

Abbreviation: H-Pro-AMC

CAS Number: 96643-94-6

Molecular Formula: C15H16N2O3

Molecular Weight: 272.30

Specifications: 100 mg / 500 mg / 1 g (Custom packaging supported)

Form: Lyophilized powder

Product Number: G12060008

Color: Blue fluorescence

Stock: In stock

H-Pro-AMC belongs to the AMC (7-amino-4-methylcoumarin) fluorescent substrate series. In the substrate molecule, L-proline is connected via an amide bond to the 7-amino group of AMC, quenching the AMC fluorophore due to acylation, rendering the substrate itself nearly non-fluorescent.

When proline-specific proteases (such as PAP, FAP, etc.) recognize the free proline residue at the N-terminus of the substrate and hydrolyze the amide bond between Pro and AMC, free AMC is released. The restoration of the free 7-amino group produces an intense blue fluorescence signal (Ex 360 nm / Em 460 nm). The fluorescence intensity is proportional to enzyme activity, enabling real-time continuous kinetic monitoring.

✅ High Sensitivity
The released AMC fluorescent signal after substrate hydrolysis is strong with low background, suitable for low-abundance enzyme activity detection and kinetic analysis.

✅ High Specificity
With proline as the recognition group, it is more suitable for proline-related peptidase systems and can be used for zymography analysis and inhibitor differentiation design.

✅ Dual Forms Available
The HBr salt form is suitable for aqueous systems, while the free base form is suitable for organic solvent systems.

✅ Multipurpose
Suitable for enzyme kinetic studies, inhibitor screening, drug development, and research on the function of tumor-related proteases.

✅ In Stock
Ample inventory allows for quick delivery to meet research and production schedule needs.

• Enzymology and Biology Research: Precisely determine prolyl aminopeptidase (PAP) activity and study its function in protein metabolism and signal transduction.

• Diagnostic Tool Development: Serves as a core raw material for research and development of detection methods and enzyme activity assay kits.

• Fibrotic Disease Research: Used for detecting proline-related peptidase activity, supporting fibrosis mechanism studies and drug screening.

Q1: What is H-Pro-AMC? Which enzymes does it detect?

A: H-Pro-AMC is an AMC fluorescent substrate that releases a highly sensitive fluorescent signal upon enzymatic cleavage. It is primarily used to detect:

• Prolyl Aminopeptidase (PAP): Specifically cleaves N-terminal proline.

• Fibroblast Activation Protein (FAP/Seprase): A key marker enzyme in the tumor microenvironment.

• Dipeptidyl Peptidase family-related enzymes (e.g., DPP-IV): Requires the use of inhibitors for differentiation.

• Prolyl Endopeptidase (PEP/PREP): Can hydrolyze this substrate under specific conditions.

Q2: What is the difference between H-Pro-AMC HBr (115388-93-7) and H-Pro-AMC (96643-94-6)?

A: The core active component is the same; the difference lies in the salt form.

• H-Pro-AMC is the free base, more hydrophobic, and must first be dissolved in DMSO or DMF.

• H-Pro-AMC HBr is the salt form, typically more convenient for preparing working solutions in aqueous buffer systems, reducing the need for organic solvents. For most biological enzyme activity assays, the HBr salt form is recommended for easier operation and to avoid potential inhibition of enzyme activity by high concentrations of organic solvents.

Q3: What are the excitation and emission wavelengths for the AMC substrate?

A: The free AMC released after enzymatic cleavage produces blue fluorescence:

• Excitation Wavelength (Ex): Approximately 360 nm~380 nm.

• Emission Wavelength (Em): Approximately 440 nm~460 nm.

Q4: How specific is this substrate? Can it differentiate between different proline enzymes?

A: H-Pro-AMC is a broad-spectrum proline N-terminal substrate; PAP, FAP, and POP (prolyl oligopeptidase) can all hydrolyze it. To differentiate specific enzyme activities (e.g., distinguish FAP from DPP-IV), it usually requires the use of specific enzyme inhibitors (such as Z-Pro-Pronal) or experimental design using different pH conditions.

Q5: How to detect fluorescence in experiments?

A: After the substrate is hydrolyzed by the enzyme, releasing AMC, real-time signals can be monitored using a fluorescence spectrophotometer or a multi-functional microplate reader with appropriate excitation/emission wavelengths. The rate of fluorescence increase is positively correlated with enzyme activity.

Q6: Is this product suitable for developing detection kits?

A: Highly suitable. This product offers high batch-to-batch stability, low background fluorescence, and excellent signal-to-noise ratio, making it an ideal core raw material for developing prolyl aminopeptidase/FAP activity detection kits, especially suitable for high-throughput drug screening kits and research-use enzyme activity assay kits.

Q7: What about product safety?

A: For research use only. Not for human use, clinical diagnosis, or animal therapy. Please wear appropriate protective equipment according to chemical safety operating procedures during experiments, and avoid direct skin/eye contact.

Q8: What packaging specifications are available? Is customization supported?

A: Three standard specifications are available: 100 mg (trial), 500 mg (regular), and 1 g (large scale). Custom packaging is supported (5 g, 10 g, 50 g, even kilogram scale) to meet different R&D or production stage needs. Custom services and custom synthesis of other amino acid-AMC substrates in the same series are also available.