As a member of the sulfatase family, arylsulfatase A is encoded by the ARSA gene and required for the lysosomal degradation of cerebroside-3-sulfate, a sphingolipid sulfate ester and a major constituent of the myelin sheet (1). ARSA deficiency results in metachromatic leukodystrophy (MLD), a lysosomal storage disease in the central and peripheral nervous systems with severe and progressive neurological symptoms (2). The deduced amino acid sequence of mouse ARSA consists of a signal peptide (residues 1-17) and a mature chain (residues 18-506) (3). Recombinant mouse ARSA corresponds to the mature chain and has sulfatase activity as described in Activity Assay Protocol.
Recombinant Mouse Arylsulfatase A/ARSA Protein, CF
R&D Systems | Catalog # 2858-SU
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Key Product Details
- R&D Systems NS0-derived Recombinant Mouse Arylsulfatase A/ARSA Protein (2858-SU)
- 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 Arylsulfatase A/ARSA protein
Ser18-Ser506, with a C-terminal 10-His tag
Ser18-Ser506, with a C-terminal 10-His tag
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
Ser18
Predicted Molecular Mass
54 kDa
SDS-PAGE
60 kDa, reducing conditions
Activity
Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS).
The specific activity is >100 pmol/min/µg, as measured under the described conditions.
The specific activity is >100 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2858-SU
| 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: Arylsulfatase A/ARSA
References
- Lukatela, G. et al. (1998) Biochemistry 37:3654.
- Parenti, G. et al. J. (1997) Curr. Opin. Genet. & Dev. 7:386.
- Kreysing, J. et al. (1994) Genomics 19:249.
Alternate Names
ARSA, Cerebroside 3-sulfatase
Gene Symbol
ARSA
UniProt
Additional Arylsulfatase A/ARSA Products
Product Documents for Recombinant Mouse Arylsulfatase A/ARSA Protein, CF
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Mouse Arylsulfatase A/ARSA Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Mouse Arylsulfatase A/ARSA Protein, CF (2858-SU):
Materials
- Assay Buffer: 50 mM Sodium Acetate, pH 5.0
- Recombinant Mouse Arylsulfatase A/ARSA (rmARSA) (Catalog # 2858-SU)
- Substrate: 4-Nitrocatechol Sulfate (PNCS) (Sigma, Catalog # N-7251), 100 mM stock in diH2O
- 0.2 M Sodium Hydroxide
- 96-well Clear Plate (Costar, Catalog # 92592)
- Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
- Dilute rmARSA to 20 µg/mL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- Combine in triplicate 75 µL of 20 µg/mL rmARSA and 75 µL of 2 mM Substrate. Include a Substrate Blank containing Assay Buffer in place of rmARSA.
- Incubate reactions for 30 minutes at 37 °C.
- Stop reactions by adding 150 µL 0.2 M sodium hydroxide to each for a final concentration of 0.1 M sodium hydroxide per reaction.
- Load into a clear 96-well plate 200 µL from each reaction vial.
- Read at 510 nm (absorbance) 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 4-Nitrocatechol (PNC) (Sigma, Catalog # N15553).
- rmARSA: 1 µg
- Substrate: 0.5 mM
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