Description
GoldBio’s N-Acetyl-3,7-dihydroxyphenoxazine (ADHP), also known as Amplex Red, is a highly sensitive fluorogenic substrate used for detecting hydrogen peroxide (H₂O₂) and monitoring oxidase activity in biochemical and cell-based assays. In the presence of horseradish peroxidase (HRP), ADHP reacts with H₂O₂ to produce the intensely fluorescent molecule resorufin, making it ideal for quantifying glucose, choline, cholesterol, and reactive oxygen species (ROS). With low background fluorescence, high stability, and excellent sensitivity in the nanomolar range, ADHP is the gold standard reagent for redox biology, metabolic profiling, and antioxidant screening. Whether you're conducting enzymatic assays or oxidative stress research, GoldBio’s ADHP delivers superior performance, reproducibility, and trusted results in every experiment.
Common Applications Using ADHP (Amplex® Red)
(Click each for more information)
Fluorometric Detection of Hydrogen Peroxide
-
Purpose: To sensitively detect H₂O₂ in biological or biochemical assays.
-
How It Works: In the presence of horseradish peroxidase (HRP), ADHP reacts with hydrogen peroxide to produce the fluorescent molecule resorufin. The fluorescence is directly proportional to H₂O₂ concentration.
-
Applications: Measurement of oxidative stress, ROS activity, or enzyme-coupled reactions that produce H₂O₂.
Zhou, M., Diwu, Z., Panchuk-Voloshina, N., & Haugland, R. P. (1997). A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. Analytical biochemistry, 253(2), 162-168.
Coupled Enzyme Assays for Monitoring Oxidase Activity
-
Purpose: To quantify activity of oxidases (e.g., glucose oxidase, choline oxidase) by coupling their enzymatic output to H₂O₂ generation..
-
How It Works: Substrate oxidation by the target enzyme generates H₂O₂, which then reacts with ADHP and HRP to form fluorescent resorufin.
-
Applications: Used to study glucose oxidase, galactose oxidase, urate oxidase, and others in enzymology.
Zhou, M., Diwu, Z., Panchuk-Voloshina, N., & Haugland, R. P. (1997). A stable nonfluorescent derivative of resorufin for the fluorometric determination of trace hydrogen peroxide: applications in detecting the activity of phagocyte NADPH oxidase and other oxidases. Analytical biochemistry, 253(2), 162-168.
Quantification of Glucose, Choline, and Cholesterol in Biofluids
-
Purpose: To measure clinically relevant metabolites using enzyme-coupled fluorescence detection.
-
How It Works: Target substrates (e.g., glucose) are oxidized enzymatically to produce H₂O₂, which is then quantified using the ADHP/HRP system.
-
Applications: Used in serum, plasma, or cell lysates for metabolic profiling, clinical diagnostics, and bioassays.
Morita, S. Y., Tsuji, T., & Terada, T. (2020). Protocols for enzymatic fluorometric assays to quantify phospholipid classes. International journal of molecular sciences, 21(3), 1032.
Screening of Antioxidants and ROS Scavengers
-
Purpose: To assess the ability of test compounds to reduce hydrogen peroxide (H₂O₂) or mitigate oxidative stress, using a sensitive fluorescence-based assay.
-
How It Works:Test compounds are incubated with H₂O₂ and ADHP/HRP. A reduction in fluorescence indicates antioxidant activity or ROS scavenging capacity.
-
Applications: CSuitable for high-throughput antioxidant screening, evaluation of mitochondrial and cellular ROS scavengers, drug and natural product discovery, and mechanistic redox biology studies.
Grivennikova, V. G., Kareyeva, A. V., & Vinogradov, A. D. (2018). Oxygen-dependence of mitochondrial ROS production as detected by Amplex Red assay. Redox biology, 17, 192-199.
Storage/Handling:
Store at 4°C
Product Specifications
ADHP
Amplex® Red
N-Acetyl-3,7-dihydroxyphenoxazine
Formula: C14H11NO4
MW: 257.24 g/mol
