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Recombinant Human Aminoacylase/ACY1 Protein, CF

Catalog #: 2900-ZN
1 Citation Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
2900-ZN-020
R&D Systems Recombinant Proteins and Enzymes
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Citations (1)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Aminoacylase/ACY1 Protein, CF Summary

Product Specifications

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.
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 conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Aminoacylase/ACY1 protein
Thr2-Ser408, with a C-terminal 10-His tag
Accession # Q03154
Accession #
Q03154
N-terminal Sequence
Analysis
Thr2
Predicted Molecular Mass
47 kDa
SDS-PAGE
42 kDa, reducing conditions

Product Datasheets

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2900-ZN

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

2900-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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 0.1 M Potassium Phosphate, pH 7.0
  • Recombinant Human Aminoacylase/ACY1 (rhACY1) (Catalog # 2900-ZN)
  • Substrate: N-Ac-Met (Sigma, Catalog # 1310), 1 M in DMSO
  • 96-well Clear UV Plate (Corning, Catalog # 3635)
  • Plate reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute Substrate to 40 mM in Assay Buffer.
  2. Load 90 µL of 40 mM Substrate to wells of the UV plate.
  3. Incubate plate at 25 °C for 10 minutes.
  4. Dilute rhACY1 to 200 µg/mL in Assay Buffer.
  5. Load 10 µL of 200 µg/mL rhACY1 to each well containing 90 µL of Substrate. Include a Substrate Blank with 90 µL of Substrate and 10 µL of Assay Buffer.
  6. Incubate plate at 25 °C for 15 minutes.
  7. Read at a wavelength of 238 nm (bottom read) in endpoint mode.
  8. 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).

Per Well:
  • rhACY1: 2 µg
  • Substrate N-Ac-Met: 36 mM
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Aminoacylase/ACY1

The human 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 as described in Activity Assay Protocol.

References
  1. Lindner, H.A. et al. (2003) J. Biol. Chem. 278:44496.
  2. Maceyka, M. et al. (2004) FEBS Lett. 568:30.
Entrez Gene IDs
95 (Human); 109652 (Mouse)
Alternate Names
ACY1; ACY-1; ACY1D; acylase; aminoacylase 1; Aminoacylase; aminoacylase-1; EC 3.5.1; EC 3.5.1.14; N-acyl-L-amino-acid amidohydrolase

Citation for Recombinant Human Aminoacylase/ACY1 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Serum aminoacylase-1 is a novel biomarker with potential prognostic utility for long-term outcome in patients with delayed graft function following renal transplantation.
    Authors: Welberry Smith, Matthew, Zougman, Alexandr, Cairns, David A, Wilson, Michelle, Wind, Tobias, Wood, Steven L, Thompson, Douglas, Messenger, Michael, Mooney, Andrew, Selby, Peter J, Lewington, Andrew J, Banks, Rosamond
    Kidney Int, 2013-06-05;84(6):1214-25.
    Species: Human
    Sample Types: Serum
    Applications: ELISA (Standard)

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