Aromatase (CYP19A) Activity Assay Kit (Fluorometric) (ab273306)
Key features and details
- Assay type: Enzyme activity (quantitative)
- Detection method: Fluorescent
- Platform: Microplate
- Sample type: Cell Lysate, Tissue Lysate
Overview
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Product name
Aromatase (CYP19A) Activity Assay Kit (Fluorometric) -
Detection method
Fluorescent -
Sample type
Cell Lysate, Tissue Lysate -
Assay type
Enzyme activity (quantitative) -
Assay duration
Multiple steps standard assay -
Product overview
Aromatase Activity Assay Kit (Fluorometric) (ab273306) enables rapid measurement of native or recombinant aromatase activity in biological samples such as placental microsomes and S9 fractions.
The assay utilizes a fluorogenic substrate that is converted into a highly fluorescent metabolite detected in the visible range (Ex/Em = 488/527 nm), ensuring a high signal-to-background ratio with little interference by autofluorescence.
A highly selective aromatase inhibitor is provided for determination of aromatase activity in heterogeneous biological samples, where other CYP isozymes may contribute to substrate metabolism. The inhibitor displays greater than 100-fold selectivity for aromatase over other enzymes, ensuring targeted inhibition. Aromatase specific activity is calculated by running parallel reactions in the presence and absence of the inhibitor and subtracting any residual activity detected with the inhibitor present.
The kit contains a complete set of reagents sufficient for 50 sets of paired reactions (in the presence and absence of inhibitor) and can detect a minimum of 25 nU aromatase activity.
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Platform
Microplate
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Beta-NADP+ Stock (100X) (Lyophilized) 1 vial Aromatase Assay Buffer 1 x 100ml Aromatase Inhibitor (Letrozole) (Lyophilized) 1 vial Aromatase Substrate (Lyophilized) 1 vial Fluorescence Standard (1 mM) 1 x 50µl NADPH Generating System (100X) (Lyophilized) 1 vial Recombinant Human Aromatase (Lyophilized) 1 vial -
Research areas
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Function
Catalyzes the formation of aromatic C18 estrogens from C19 androgens. -
Tissue specificity
Brain, placenta and gonads. -
Involvement in disease
Defects in CYP19A1 are a cause of aromatase excess syndrome (AEXS) [MIM:139300]; also known as familial gynecomastia. AEXS is characterized by an estrogen excess due to an increased aromatase activity.
Defects in CYP19A1 are the cause of aromatase deficiency (AROD) [MIM:107910]. AROD is a rare disease in which fetal androgens are not converted into estrogens due to placental aromatase deficiency. Thus, pregnant women exhibit a hirsutism, which spontaneously resolves after post-partum. At birth, female babies present with pseudohermaphroditism due to virilization of extern genital organs. In adult females, manifestations include delay of puberty, breast hypoplasia and primary amenorrhoea with multicystic ovaries. -
Sequence similarities
Belongs to the cytochrome P450 family. -
Cellular localization
Membrane. - Information by UniProt
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Alternative names
- ARO
- ARO1
- Aromatase
see all
Images
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Example standard curve
Standard curve of Aromatase Substrate metabolite fluorescence. One mole of fluorescence standard corresponds to the metabolism of one mole of Aromatase Substrate.
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Reaction KineticsReaction kinetics of fluorogenic substrate metabolism in donor-pooled human placental microsomes (0.5 mg/mL) at 37°C in the presence and absence of the competitive aromatase inhibitor letrozole (the no inhibitor condition contained assay buffer with 0.2% acetonitrile as a solvent control).
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Specific activity of aromatase in human placental microsome samplesSpecific activity of aromatase in human placental microsome samples (mean ± SEM of four independent replicates). Assays were performed according to the kit protocol.