Cathepsin A/lyososomal carboxypeptidase A is a member of the serine carboxypeptidase family (1). Cathepsin A is a multifunctional enzyme that expresses deaminidase and esterase activities at neutral pH and carboxypeptidase activity at acidic pH. Also known as protective protein, its association with beta -galactosidase ( beta -gal) and neuraminidase is essential for beta -gal stability and neuraminidase activation in the lysosomes. Inherited deficiency of Cathepsin A causes the lysosomal storage disorder galactosialidosis, characterized by a combined secondary deficiency of beta -gal and neuraminidase. Cathepsin A is capable of hydrolyzing a variety of bioactive peptide hormones including tachykinins, indicating that extralysosomal Cathepsin A plays a role in regulation of functions of these molecules (2). Cathepsin A is synthesized as a single-chain precursor and processed into heavy (32 kDa) and light (20 kDa) chains, which are linked by disulfide bonds.
Recombinant Human Cathepsin A/Lysosom Carboxypeptidase A, CF
R&D Systems | Catalog # 1049-SE
Key Product Details
- R&D Systems NS0-derived Recombinant Human Cathepsin A/Lysosom Carboxypeptidase A (1049-SE)
- 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
Accession Number
Structure / Form
Applications
Product Specifications
Source
Ala29-Tyr480, with a C-terminal 10-His tag
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity is >75 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
1049-SE
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| 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: Cathepsin A/Lysosomal Carboxypeptidase A
References
- Pshezhetsky, A.V. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) p. 1923, Academic Press, San Diego.
- Hiraiwa, M. (1999) Cell. Mol. Life. Sci. 56:894.
Alternate Names
Gene Symbol
UniProt
Additional Cathepsin A/Lysosomal Carboxypeptidase A Products
- All Products for Cathepsin A/Lysosomal Carboxypeptidase A
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- Cathepsin A/Lysosomal Carboxypeptidase A ELISA Kits
- Cathepsin A/Lysosomal Carboxypeptidase A Lysates
- Cathepsin A/Lysosomal Carboxypeptidase A Primary Antibodies
- Cathepsin A/Lysosomal Carboxypeptidase A Proteins and Enzymes
Product Documents for Recombinant Human Cathepsin A/Lysosom Carboxypeptidase A, 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 Cathepsin A/Lysosom Carboxypeptidase A, CF
For research use only
Related Research Areas
Citations for Recombinant Human Cathepsin A/Lysosom Carboxypeptidase A, CF
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Protocols
View specific protocols for Recombinant Human Cathepsin A/Lysosom Carboxypeptidase A, CF (1049-SE):
- Activation Buffer: 25 mM MES, 5 mM DTT, pH 6.0
- Assay Buffer: 25 mM MES, 5 mM DTT, pH 5.5
- Recombinant Human Cathepsin A/Lysosomal Carboxypeptidase A (rhCathepsin A) (Catalog # 1049-SE)
- Recombinant Human Cathepsin L (rhCathepsin L) (Catalog # 952-CY)
- E 64 (Tocris Catalog # 5208) , 50 mM stock in DMSO
- Substrate: MCA-Arg-Pro-Pro-Gly-Phe-Ser-Ala-Phe-Lys(DNP)-OH (Catalog # ES005)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhCathepsin A to 100 µg/mL in Activation Buffer.
- Dilute rhCathepsin L to 10 µg/mL in Activation Buffer.
- Combine equal volumes of rhCathepsin A and rhCathepsin L for final concentrations of 50 µg/mL and 5 µg/mL, respectively.
- Incubate reaction at 37 °C for 30 minutes.
- Stop reaction by adding E-64 to a final concentration of 10 µM.
- Dilute activated rhCathepsin A to 2.0 ng/µL in Assay Buffer.
- Dilute Substrate to 20 µM in Assay Buffer.
- Load 50 µL of 2.0 ng/µL rhCathepsin A into a plate, and start the reaction by adding 50 µL of 20 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 20 µM Substrate.
- Read at excitation and emission wavelengths of 320 nm and 405 nm (top read), respectively, in kinetic mode for 5 minutes.
- 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 MCA-Pro-Leu-OH (Bachem, Catalog # M-1975)
Per Well:
- rhCathepsin A: 0.1 µg
- Substrate: 10 µM
FAQs for Recombinant Human Cathepsin A/Lysosom Carboxypeptidase A, CF
-
Q: Where does Cathepsin A cleave Mca-RPPGFSAFK(Dnp)-OH Fluorogenic Peptide Substrate, Catalog # ES005?
A: Although the QC assay conditions provided on our recombinant Cathepsin A datasheets should favor carboxypeptidase activity, it is possible that there is more than one site in the ES005 peptide recognized and cleaved by Cathepsin A. We have not performed verification experiments to confirm the cleavage site preferred in our reaction conditions.