4-Methylumbelliferyl phosphate – Research Use Only

4-Methylumbelliferyl phosphate (4-MUP) is a highly sensitive fluorogenic substrate supplied as a 5 g powder (Catalog #B2014906, MW 256.15 g/mol). Upon hydrolysis by phosphatases (alkaline phosphatase, acid phosphatase, etc.), it releases the highly fluorescent 4-methylumbelliferone (4-MU), enabling sensitive detection and quantification of phosphatase activity in biochemical, clinical, and high-throughput screening assays.

Scientific Overview

4-Methylumbelliferyl phosphate (4-MUP) is a non-fluorescent substrate that is hydrolyzed by phosphatases to release the highly fluorescent product 4-methylumbelliferone (4-MU, excitation ~360 nm, emission ~450 nm). This fluorogenic reaction provides excellent sensitivity and a wide dynamic range, making 4-MUP the preferred substrate for detecting alkaline phosphatase, acid phosphatase, and other phosphatase activities in ELISA, high-throughput screening, kinetic assays, and histochemical applications.

This 4-Methylumbelliferyl phosphate is suitable for:

• Fluorometric detection and quantification of phosphatase activity
• High-throughput enzyme screening and kinetic studies
• ELISA and immunoassay development (as alkaline phosphatase substrate)
• Biochemical characterization of protein phosphatases
• Histochemical and cytochemical localization of phosphatase activity

Usage & Handling Guidance

Store the powder at -20 °C. Reconstitute in ultrapure water or assay buffer to the desired concentration (typically 0.1–1 mM for most assays). Prepare fresh working solutions as needed. The reaction is usually performed at pH 8–10 for alkaline phosphatase or pH 4–6 for acid phosphatase, with fluorescence measured at Ex/Em ≈ 360/450 nm.

• Recommended applications: Phosphatase assays and ELISA
• Working concentration: 0.1–1 mM final concentration (optimize for your enzyme and instrument)
• Stability: Powder is stable at -20 °C; aqueous solutions should be used fresh or stored frozen
• Handling: Protect from light; use black plates or low-fluorescence consumables for maximum sensitivity

What You Get

• 5 g 4-Methylumbelliferyl phosphate as a high-purity powder
• Fluorogenic substrate (MW 256.15 g/mol) for sensitive phosphatase detection
• Biotechnology-grade material suitable for high-throughput and quantitative assays
• Large quantity ideal for method development and routine laboratory use
• For research use only (RUO)

Why Researchers Choose It

• Highly sensitive fluorogenic substrate for phosphatase activity detection
• Excellent dynamic range and low background in fluorescence-based assays
• Preferred substrate for alkaline phosphatase in ELISA and high-throughput screening
• High-purity powder format ensures reproducible performance
• Cost-effective bulk size for routine laboratory and method development use

Frequently Asked Questions (FAQ)

• What enzyme does 4-MUP detect?
  It is hydrolyzed by alkaline phosphatase, acid phosphatase, and many other phosphatases, releasing fluorescent 4-methylumbelliferone.
• What are the excitation/emission wavelengths?
  The product 4-MU is typically measured at excitation ~360 nm and emission ~450 nm.
• How do I prepare the substrate solution?
  Dissolve the powder in ultrapure water or assay buffer to 0.1–1 mM. Prepare fresh or store aliquots at -20 °C.
• Is this suitable for ELISA?
  Yes. 4-MUP is one of the most sensitive substrates for alkaline phosphatase-based ELISA detection.
• How should I store the product?
  Store the powder at -20 °C for long-term stability.

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References

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