The mouse ACY1 gene encodes aminoacylase, a member of the M20 family of metalloproteases (1). ACY1 plays a general role in the cytosolic breakdown of acetylated amino acids generated during protein degradation. It also interacts with sphingosine kinase type 1, which is involved in promoting cell growth and inhibiting apoptosis of tumor cells (2). The full-length protein is expressed and the purified enzyme is active in the assay described in the Activity Assay Protocol.
Recombinant Mouse Aminoacylase/ACY1 Protein, CF
R&D Systems | Catalog # 2700-ZN
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Aminoacylase/ACY1 Protein (2700-ZN)
- 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
NS0
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived mouse Aminoacylase/ACY1 protein
Met1-Ser408, with a C-terminal 10-His tag
Accession # NP_079647
Met1-Ser408, with a C-terminal 10-His tag
Accession # NP_079647
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
No results obtained: Gln18 predicted
Predicted Molecular Mass
45 kDa
SDS-PAGE
42 kDa, reducing conditions
Activity
Measured by its ability to cleave N-acetyl-L-Methione (Ac-Met).
The specific activity is >4,000 pmol/min/µg, as measured under the described protocol.
The specific activity is >4,000 pmol/min/µg, as measured under the described protocol.
Formulation, Preparation, and Storage
2700-ZN
| 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: Aminoacylase/ACY1
References
- Lindner, H.A. et al. (2003) J. Biol. Chem. 278:44496.
- Maceyka, M. et al. (2004) FEBS Lett. 568:30.
Alternate Names
ACY1
Gene Symbol
ACY1
UniProt
Additional Aminoacylase/ACY1 Products
Product Documents for Recombinant Mouse Aminoacylase/ACY1 Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Aminoacylase/ACY1 Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Aminoacylase/ACY1 Protein, CF (2700-ZN):
Materials
- Assay Buffer: 0.1 M Potassium Phosphate, pH 7.0
- Recombinant Mouse Aminoacylase/ACY1 (rmACY1) (Catalog # 2700-ZN)
- Substrate: N-Acetyl-L-Methionine (N-Ac-Met) (Sigma, Catalog # 1310), 1 M stock in DMSO
- 96 well Clear UV Plate (Corning, Catalog # 3635)
- Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute Substrate to 40 mM in Assay Buffer.
- In a plate load 90 µL of 40 mM Substrate to wells.
- Incubate plate at 25 °C for 10 minutes.
- Dilute rmACY1 to 200 µg/mL in Assay Buffer.
- Load 10 µL of 200 µg/mL rmACY1 to each well containing 90 µL of Substrate. Include a Substrate Blank with 90 µL of 40 mM Substrate and 10 µL of Assay Buffer.
- Incubate plate at 25 °C for 15 minutes.
- Read at a wavelength of 238 nm (bottom read) in endpoint mode.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Abs* (OD) x Conversion Factor** (pmol/OD) |
| Incubation time (min) x amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard N-Acetyl-L-Methionine (Sigma, Catalog # 1310).
- rmACY1: 2 µg
- Substrate: 36 mM
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