Amiloride binding protein (ABP1), also known as diamine oxidase (DAO) and amine oxidase copper domain-containing protein 1 (AOC1), is an approximately 110 kDa member of the copper-containing amine oxidase family (1, 2). ABP1 is a glycosylated enzyme that forms disulfide-linked homodimers (3). It is stored in cytoplasmic granules of epithelial cells in the kidney, placenta, uterus, lung, and intestine (1, 4), and its extracellular release can be induced by heparin (5). ABP1 exhibits a substrate preference for histamine and putrescine, generating hydrogen peroxide and ammonia in an oxidative deamination reaction (3). ABP1 activity plays an important role in the catabolism of histamine and other bioactive polyamines (6, 7). Deficiencies in ABP1 can result in histaminosis or dietary histamine intolerance (8, 9).
Recombinant Human ABP1/AOC1 Protein, CF
R&D Systems | Catalog # 8298-AO
Loading...
Key Product Details
- R&D Systems Sf 21 (baculovirus)-derived Recombinant Human ABP1/AOC1 Protein (8298-AO)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
Sf 21 (baculovirus)
Accession Number
Applications
Enzyme Activity
Loading...
Product Specifications
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human ABP1/AOC1 protein
Glu20-Val751, with a N-terminal 6-His tag
Glu20-Val751, with a N-terminal 6-His tag
Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Inconclusive, His predicted. Protein identity confirmed by MS analysis of tryptic fragments.
Predicted Molecular Mass
84 kDa
SDS-PAGE
83-92 kDa, reducing conditions
Activity
Measured by its ability to produce hydrogen peroxide during the oxidation of histamine.
The specific activity is >40 pmol/min/μg, as measured under the described conditions.
The specific activity is >40 pmol/min/μg, as measured under the described conditions.
Formulation, Preparation, and Storage
8298-AO
| Formulation | Supplied as a 0.2 μm filtered solution in HEPES, NaCl, Glycerol. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: ABP1/AOC1
References
- Novotny, W.F. et al. (1994) J. Biol. Chem. 269:9921.
- Barbry, P. et al. (1990) Proc. Natl. Acad. Sci. USA 87:7347.
- McGrath, A.P. et al. (2009) Biochemistry 48:9810.
- Liang, X.H. et al. (2010) Endocrinology 151:5007.
- Klocker, J. et al. (2004) Vascul. Pharmacol. 40:293.
- Jones, B.L. and G.L. Kearns (2011) Clin. Pharmacol. Ther. 89:189.
- Bieganski, T. et al. (1983) Biochim. Biophys. Acta 756:196.
- Sattler, J. and W. Lorenz (1990) J. Neural Transm. Suppl. 32:291.
- Maintz, L. and N. Novak (2007) Am. J. Clin. Nutr. 85:1185.
Long Name
Amiloride Binding Protein 1
Alternate Names
AOC1, Diamine Oxidase, Histaminase, KAO
Entrez Gene IDs
26 (Human)
Gene Symbol
AOC1
UniProt
Additional ABP1/AOC1 Products
Product Documents for Recombinant Human ABP1/AOC1 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human ABP1/AOC1 Protein, CF
For research use only
Related Research Areas
Citations for Recombinant Human ABP1/AOC1 Protein, CF
Powered by Bioz
Customer Reviews for Recombinant Human ABP1/AOC1 Protein, CF
There are currently no reviews for this product. Be the first to review Recombinant Human ABP1/AOC1 Protein, CF and earn rewards!
Have you used Recombinant Human ABP1/AOC1 Protein, CF?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Protocols
View specific protocols for Recombinant Human ABP1/AOC1 Protein, CF (8298-AO):
Materials
- Assay Buffer: 50 mM HEPES, pH 7.5
- Recombinant Human ABP1/AOC1 (rhABP1) (Catalog # 8298-AO)
- Coupling Enzyme: Horseradish Peroxidase (HRP) (250-330 U/mg) (Sigma, Catalog # P8375), 250 units/mL stock in 0.1 M Sodium Phosphate, pH 8.0
- Substrate Component 1: Histamine (Sigma, Catalog # H7250), 20 mM stock in deionized water
- Substrate Component 2: Amplex Ultra Red (AUR) (Molecular Probes, Catalog # A36006), 10 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhABP1 to 1 ng/µL in Assay Buffer.
- Prepare a Substrate Mixture containing 10 µM Histamine, 2 units/mL HRP, and 100 µM AUR in Assay Buffer.
- Load 50 µL of 1 ng/µL rhABP1 into wells of a plate and start the reactions by adding 50 μL of Substrate Mixture. Include a Substrate Blank containing Assay Buffer instead of rhABP1.
- Read at excitation and emission wavelengths of 544 nm and 590 nm (top read), respectively in kinetic mode. Note: A cutoff must be set manually at a wavelength of 570 nm.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using a fluorescent standard prepared by incubating 50 µM AUR, 1 unit/mL HRP, 5 µM Histamine, and a curve of Hydrogen Peroxide (Sigma, Catalog # H1009) in Assay Buffer. Use this oxidized AUR curve to determine the conversion factor.
- rhABP1: 0.05 µg
- Histamine: 5 µM
- HRP: 1 unit/mL
- AUR: 50 µM
Loading...