Recombinant Mouse Aminoacylase/ACY1 Protein, CF Summary
Met1-Ser408, with a C-terminal 10-His tag
Accession # NP_079647
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CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|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.
- 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
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.
- Lindner, H.A. et al. (2003) J. Biol. Chem. 278:44496.
- Maceyka, M. et al. (2004) FEBS Lett. 568:30.
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