Aminopeptidase A Protein, Human (sf9, His)
Based on 1 Customer Validation
Aminopeptidase A protein has calcium-regulating activity and plays a key role in central hypertension by preferentially cleaving N-terminal acidic residues, including angiotensin II. The specificity of this enzyme highlights its involvement in the regulation of blood pressure-related peptides, suggesting potential implications for the physiological mechanisms of hypertension. Aminopeptidase A Protein, Human (sf9, His) is the recombinant human-derived Aminopeptidase A protein, expressed by Sf9 insect cells , with N-His labeled tag.
- Species: Human
- 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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Biological Activity
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Technical Parameters
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Product Properties
- Documentation
- Help & FAQs
Biological Activity
Description
Aminopeptidase A protein has calcium-regulating activity and plays a key role in central hypertension by preferentially cleaving N-terminal acidic residues, including angiotensin II. The specificity of this enzyme highlights its involvement in the regulation of blood pressure-related peptides, suggesting potential implications for the physiological mechanisms of hypertension. Aminopeptidase A Protein, Human (sf9, His) is the recombinant human-derived Aminopeptidase A protein, expressed by Sf9 insect cells , with N-His labeled tag.
Background
The Aminopeptidase A protein serves a pivotal role in the regulation of central hypertension, demonstrating a calcium-modulated preference for cleaving N-terminal acidic residues from peptides, including angiotensin II. This enzymatic activity highlights its involvement in the modulation of peptides associated with blood pressure regulation, suggesting a potential impact on the physiological mechanisms underlying hypertension. The calcium-dependent specificity of Aminopeptidase A underscores its intricate regulatory role in the processing of bioactive peptides, emphasizing its significance in cardiovascular function and potential relevance in the context of hypertensive conditions.
Verified Bioactivity
Measured by its ability to cleave the fluorogenic peptide substrate, Glu-7-amido-4-methyl coumarin (Glu-AMC) and the specific activity is >2000 pmol/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 CaCl?, 0.2 M NaCl, pH 8.0
Aminopeptidase A Protein, Human (sf9, His) (HY-P72824)
Substrate: Glu-7-amido-4-methylcoumarin
Standard: 7-amido-4-methylcoumarin
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 concentration of the standard 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 Human 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 Human Aminopeptidase A and 200 μM substrate to the wells of 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 enzyme 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:
Human Aminopeptidase A: 0.02 μg
Substrate: 100 μM
Technical Parameters
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Species Human
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Source Sf9 insect cells
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Tag N-His
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Accession
Q07075/NP_001968.3 (R41-G957)
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Molecular Construction
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N-term
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His
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ENPEP (R41-G957)
Accession # Q07075/NP_001968.3 -
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
RSCDSSGDGGPGTAPAPSHLPSSTASPSGPPAQDQDICPASEDESGQWKNFRLPDFVNPVHYDLHVKPLLEEDTYTGTVSISINLSAPTRYLWLHLRETRITRLPELKRPSGDQVQVRRCFEYKKQEYVVVEAEEELTPSSGDGLYLLTMEFAGWLNGSLVGFYRTTYTENGQVKSIVATDHEPTDARKSFPCFDEPNKKATYTISITHPKEYGALSNMPVAKEESVDDKWTRTTFEKSVPMSTYLVCFAVHQFDSVKRISNSGKPLTIYVQPEQKHTAEYAANITKSVFDYFEEYFAMNYSLPKLDKIAIPDFGTGAMENWGLITYRETNLLYDPKESASSNQQRVATVVAHELVHQWFGNIVTMDWWEDLWLNEGFASFFEFLGVNHAETDWQMRDQMLLEDVLPVQEDDSLMSSHPIIVTVTTPDEITSVFDGISYSKGSSILRMLEDWIKPENFQKGCQMYLEKYQFKNAKTSDFWAALEEASRLPVKEVMDTWTRQMGYPVLNVNGVKNITQKRFLLDPRANPSQPPSDLGYTWNIPVKWTEDNITSSVLFNRSEKEGITLNSSNPSGNAFLKINPDHIGFYRVNYEVATWDSIATALSLNHKTFSSADRASLIDDAFALARAQLLDYKVALNLTKYLKREENFLPWQRVISAVTYIISMFEDDKELYPMIEEYFQGQVKPIADSLGWNDAGDHVTKLLRSSVLGFACKMGDREALNNASSLFEQWLNGTVSLPVNLRLLVYRYGMQNSGNEISWNYTLEQYQKTSLAQEKEKLLYGLASVKNVTLLSRYLDLLKDTNLIKTQDVFTVIRYISYNSYGKNMAWNWIQLNWDYLVNRYTLNNRNLGRIVTIAEPFNTELQLWQMESFFAKYPQAGAGEKPREQVLETVKNNIEWLKQHRNTIREWFFNLLESG
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Predicted Molecular Mass
107.2 kDa
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Molecular Weight
Approximately 116 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, 10 % glycerol, 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 (241 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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