ЭЛЬФ 97

Description

ELF 97 (ELF 97 phosphate) is a synthetic fluorogenic substrate of phosphatases, aryl phosphate of the quinazolinone series. When hydrolyzed by an alkaline or acid phosphatase, the phosphate group is cleaved and the water-insoluble product ELF 97 alcohol is released, which forms a bright yellow-green, photostable fluorescent precipitate directly at the site of enzymatic activity (excitation at 345 nm, emission at 530 nm). The substrate does not have cell permeability and does not diffuse from the point of product formation, which ensures high spatial accuracy of localization of the phosphatase activity signal in cells and tissues.

Basic Attributes

• CAS No: 147394-94-3
• Chemical formula: C14H9Cl2N2O5P
• Molecular Weight: 387.109
• Exact weight: 387.11
• DSSTox ID: DTXSID10163695
• PSA Specifications: 108
• XLogP3: 1.9

Physico-chemical characteristics

• Density: 1.78±0.1 g/cm3 (predicted value)
• Boiling point: 654.9±65.0 °C (predicted value)

Chemical identification

Chemical class: aryl phosphate, a representative of the group of quinazolinones and monochlorobenzenes. Systematic name: 6-chloro-2-[5-chloro-2-(phosphonooxyl)phenyl]-4(3H)-quinazolinone (2-(5′-chloro-2′-phosphophenyl)-6-chloro-4(3H)-quinazolinone). Synonyms: ELF 97 phosphate.

Scope of application

Used in immunohistochemistry, in situ hybridization with fluorescence detection (FISH) for signal amplification, as well as for detection of endogenous alkaline phosphatase activity in intact cells by flow cytometry. It is used to study the activity of extracellular phosphatases of algae and microorganisms in natural waters - the enzymatic activity of individual cells is assessed by the intensity and localization of the fluorescent precipitate. It is in demand as a reagent that produces a bright and photostable signal that is resistant to fading during long-term microscopy compared to soluble fluorogenic substrates.

Advantages of the method

Unlike classical soluble chromogenic phosphatase substrates (e.g. BCIP/NBT or para-nitrophenyl phosphate), the hydrolysis product of ELF 97 does not diffuse from the site of the enzymatic reaction, but is deposited directly there, providing precise subcellular localization of the signal. The high photostability of the precipitate allows long-term microscopic imaging without noticeable fluorescence fading, which is critical for quantitative comparison of different samples in one experiment.

Storage and transportation

Store in an airtight container, protected from light, at the recommended storage temperature for fluorescence microscopy reagents—usually a refrigerator, away from heat and direct light, to avoid premature decomposition of the phosphate substrate.

Advantages over classical substrates

Unlike soluble chromogenic phosphatase substrates (eg BCIP/NBT or p-nitrophenyl phosphate), the hydrolysis product of ELF 97 is insoluble and precipitates directly at the point of the enzymatic reaction without diffusing into the sample environment. This is fundamentally important for the quantitative localization of enzyme activity at the subcellular level using fluorescence microscopy methods - the researcher receives an accurate topography of the signal, and not a blurry cloud of the reaction product, as when using soluble analogues. An additional advantage is the high photostability of the precipitate, which allows long-term recording without noticeable signal fading, which is especially valuable for quantitative comparison of samples.