Diamine Oxidase Assay Kit (ab241004)
Key features and details
- Detection method: Fluorescent
- Platform: Microplate reader
Overview
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Product name
Diamine Oxidase Assay Kit -
Detection method
Fluorescent -
Product overview
Diamine Oxidase Assay Kit (ab241004) provides a straightforward method to determine DAO activity of tissue lysates as well as recombinant enzymes, with a detection limit of less than 1 pmol/min of activity.
In the assay, DAO converts the provided substrate, yielding an intermediate and H2O2. H2O2 is then utilized by the DAO Enzyme Mix to generate fluorescence (Ex/Em = 535/587 nm) from the DAO Probe.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests DAO Assay Buffer 1 x 25ml DAO Enzyme Mix (Lyophilized) 1 vial DAO Positive Control (Lyophilized) 1 vial DAO Probe (in DMSO) 1 x 200µl DAO Substrate (Lyophilized) 1 vial H2O2 Standard (0.88 M) 1 x 100µl -
Relevance
Diamine Oxidase (DAO, E.C. 1.4.3.22) is an important regulator of polyamine levels in human physiology. DAO metabolizes the oxidative conversion of biogenic polyamines such as putrescine and cadaverine into aldehydes. Biogenic amines are known regulators of important biological pathways such as cell growth and signal transduction, placing DAO in an important regulatory position in maintaining cell viability. Putrescine itself is also an activator of c-jun and c-fos expression, further linking this diamine with potential roles in cancer development. DAO is notably involved in regulating growth and differentiation of intestinal mucosa, bone marrow, and other rapidly proliferating tissue. In addition to its role in tissue growth, DAO activity is important in metabolism. Loss of diamine oxidase activity can lead to histamine intolerance and histamine poisoning.
Images
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H2O2 standard curve
H2O2 standard curve
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Kinetic measurement of DAO activity from Positive ControlKinetic measurement of DAO activity from Positive Control.
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Activity determination of intestinal tissue lysate.Activity determination of intestinal tissue lysate. For this experiment, 100 mg rat intestine was used, following Diamine Oxidase Activity Assay Kit protocol and including protease inhibitor in the lysis. Lysate (7 µg) was assayed and specific activity was determined to be 1.30 nmol/min/mg lysate.