Aminopeptidase A Protein, Mouse (sf9, His)
Based on 1 Customer Validation
The aminopeptidase A protein is a key player in the regulation of central hypertension, exerting its effects by selectively cleaving the N-terminal acidic residues of peptides such as angiotensin II. The enzyme's preference for specific substrates is regulated by calcium levels, underscoring its critical role in controlling blood pressure and maintaining cardiovascular health. Aminopeptidase A Protein, Mouse (sf9, His) is the recombinant mouse-derived Aminopeptidase A protein, expressed by Sf9 insect cells , with N-His labeled tag.
- Species: Mouse
- Source: Sf9 insect cells
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Storage:Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.
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Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
Biological Activity
Description
The aminopeptidase A protein is a key player in the regulation of central hypertension, exerting its effects by selectively cleaving the N-terminal acidic residues of peptides such as angiotensin II. The enzyme's preference for specific substrates is regulated by calcium levels, underscoring its critical role in controlling blood pressure and maintaining cardiovascular health. Aminopeptidase A Protein, Mouse (sf9, His) is the recombinant mouse-derived Aminopeptidase A protein, expressed by Sf9 insect cells , with N-His labeled tag.
Background
Aminopeptidase A protein is a key player in the regulation of central hypertension, exerting its influence by selectively cleaving N-terminal acidic residues from peptides such as angiotensin II. The enzyme's preference for specific substrates is modulated by calcium levels, underscoring its crucial role in controlling blood pressure and maintaining cardiovascular health. By cleaving peptides, aminopeptidase A protein affects the balance of vasoactive substances and ultimately contributes to the regulation of vascular tone and blood pressure. Understanding the intricate mechanisms and functional implications of this protein can provide valuable insights into the development of targeted therapies for hypertension-related conditions.
Verified Bioactivity
Measured by its ability to cleave the fluorogenic peptide substrate, Glu-7-amido-4-methyl coumarin and the specific activity is > 200 pmoles/min/μg.
MCE Validation Data
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Purity - SDS-PAGE
Purity - SDS-PAGE
Assay Procedure
Materials
Assay buffer: 25 mM Tris, 50 mM CaCl2, 0.2 M NaCl, pH 8.0
Aminopeptidase A Protein, Mouse (sf9, His) (HY-P72825)
Substrate: Glu-7-amido-4-methylcoumarin
Standard: 7-amido-4-methylcoumarin (HY-D0027)
Black 96-well Plate
Fluorescence Microplate Reader
Procedure
1. Dilute 7-amido-4-methylcoumarin in assay buffer to concentrations of 0, 0.2, 0.39, 0.78, 1.56, 3.13, 6.25, 12.5, 25, 50 μmol/L. Add 100 μL of each concentration to the wells to initiate the reaction, with three replicates per concentration. Read the fluorescence in kinetic mode for 5 minutes with excitation and emission wavelengths of 380 nm and 460 nm (top read) . Plot the standard concentrations on the x-axis and the measured RFU on the y-axis to create the standard curve and obtain the standard curve equation.
2. Dilute Mouse Aminopeptidase A to 0.4 μg/mL in assay buffer.
3. Dilute the substrate to 200 μM in assay buffer.
4. Add 100 μL of 0.4 μg/mL Mouse Aminopeptidase A and 200 μM substrate to the black 96-well plate. Include a control with assay buffer and substrate only. Perform three replicates for each condition.
5. Read the RFU values in kinetic mode for 5 minutes with excitation and emission wavelengths of 380 nm and 460 nm, respectively.
6. 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 calibration standard
Per Well:
Mouse Aminopeptidase A: 0.02 μg
Substrate: 100 μM
Technical Parameters
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Species Mouse
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Source Sf9 insect cells
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Tag N-His
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Accession
P16406 (R41-P945)
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Molecular Construction
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N-term
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His
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ENPEP (R41-P945)
Accession # P16406 -
C-term
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Protein Length
Extracellular Domain
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Synonyms
ENPEP; EAP; Glutamyl Aminopeptidase; Glutamyl Aminopeptidase (Aminopeptidase A); CD249; Alternative Protein ENPEP; Gp160; Aminopeptidase; Differentiation Antigen Gp160; CD249 Antigen; Aminopeptidase A; APA; AP-A
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AA Sequence
RSCEQDTTPAPSQPPPEASTALPPQDQNVCPDSEDESGEWKNFRLPDFINPVHYDLEVKALMEEDRYTGIVTISVNLSKPTRDLWLHIRETKITKLPELRRPSGEQVPIRRCFEYKKQEYVVIQAAEDLAATSGDSVYRLTMEFKGWLNGSLVGFYKTTYMEDGQIRSIAATDHEPTDARKSFPCFDEPNKKSTYSISIIHPKEYSALSNMPEEKSEMVDDNWKKTTFVKSVPMSTYLVCFAVHRFTAIERKSRSGKPLKVYVQPNQKETAEYAANITQAVFDYFEDYFAMEYALPKLDKIAIPDFGTGAMENWGLVTYRETNLLYDPLLSASSNQQRVASVVAHELVHQWFGNTVTMDWWDDLWLNEGFASFFEFLGVNHAEKDWQMLSQVLLEDVFPVQEDDSLMSSHPVVVTVSTPAEITSVFDGISYSKGASILRMLQDWITPEKFQKGCQIYLKKFQFANAKTSDFWDSLQEASNLPVKEVMDTWTSQMGYPVVTVSGRQNITQKRFLLDSKADPSQPPSELGYTWNIPVRWADNDNSRITVYNRLDKGGITLNANLSGDAFLKINPDHIGFYRVNYEGGTWDWIAEALSSNHTRFSAADRSSFIDDAFALARAQLLNYKIALNLTMYLKSEEDFLPWERVISSVSYIISMFEDDRELYPMIETYFQGQVKPVADLLGWQDTGSHITKLLRASILGFACKMGDREALGNASQLFDSWLKGSASIPVNLRLLVYRYGMQNSGNEAAWNYTLEQYQKTSLAQEKEKLLYGLASVKDVKLLARYLEMLKDPNIIKTQDVFTVIRYISYNSYGKTMAWNWIQLNWDYLVSRFTINDRYLGRIVTIAEPFNTELQLWQMQSFFAKYPNAGAGAKPREQVLETVKNNIEWLNVNRQSIREWFASLP
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Predicted Molecular Mass
106 kDa
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Molecular Weight
Approximately 113 kDa, based on SDS-PAGE under reducing conditions.
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Glycosylation
Yes
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Purity
≥ 90%, as determined by reducing SDS-PAGE.
Product Properties
Solution
Supplied as a 0.22 μm filtered solution of 20 mM Tris, 500 mM NaCl, pH 7.4.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.
<1 EU/μg, determined by LAL method.
Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.
Shipping with dry ice.
Documentation
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Data Sheet (240 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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