BOC Sciences is committed to providing customers with high-quality fluorescent enzyme substrates.
Fluorescent enzyme substrates are substances that produce fluorescence after the action of specific enzymes. They have no fluorescence effect and can be used to detect enzyme activities in living cell systems and solutions. The fluorescent enzyme substrates we provide are quite stable, can be stored for a long time, and can react with various enzymes.
Enzyme-Linked Immunosorbent Assay is the most widely used solid-phase immunoassay method, and it has extraordinary applicability in basic research and clinical trials. Its principle is to let the antibody bind to the enzyme complex and then detect it by color development. After adding the substrate of the enzyme reaction, the substrate is catalyzed by the enzyme to become a colored product. Commonly used chromogenic substrates are o-phenylenediamine, p-nitrophenyl-phosphoric acid, and o-nitrophenyl-β-Dgal actopyranoside.
Later, the enzyme-linked reaction began to pursue higher sensitivity and flexibility. Fluorescent substrates are developed after chromogenic substrates.
Introduction of Fluorescent Enzyme Substrates
Compared with chromogenic substrates, the detection sensitivity of fluorescent substrates is higher, and several fluorescent materials of different colors can be used at the same time. When the substrate is hydrolyzed by a specific enzyme, the fluorescent enzyme substrates show a linear increase in fluorescence intensity proportional to the enzyme activity. Fluorescence microscope, flow cytometer, and fluorescence microplate reader can detect fluorescent enzyme substrates. The fluorescent technology using fluorescent enzyme substrates is simple to operate, low in cost, and high in sensitivity.
Examples of Fluorescent Enzyme Substrates
The substrates of β-galactosidase (β-gal) include 4-methylumbelliferyl-β-D-galactopyranoside, fluorescein di-β-D-galactosidase, 4-methylumbelliferyl-galactoside 6-sulfate, etc. 4-methylum belliferyl-β-D-galactopyranoside is decomposed into 4-Methylumbelliferone by the action of β-gal. The latter can emit fluorescence, the excitation wavelength is 360nm, and the emission wavelength is 450nm. Also, the substrate of alkaline phosphatase (ALP) is 4-methylumbelliferyl phosphate. The substrates of horseradish peroxidase (HRP) include p-hydroxyphenylacetic acid and 3-(p-hydroxyphenyl) propionic acid.