Recombinant Human Arylsulfatase A/ARSA Protein, CF

R&D Systems | Catalog # 2485-SU

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human Arylsulfatase A/ARSA Protein (2485-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

AAH14210

Applications

Enzyme Activity
View all Arylsulfatase A/ARSA Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human Arylsulfatase A/ARSA protein
Arg19-Ala507, 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

Arg19

Predicted Molecular Mass

53 kDa

SDS-PAGE

63 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS).
The specific activity is >25 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

2485-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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Arylsulfatase A/ARSA

As a member of the sulfatase family, ARSA is required for the lysosomal degradation of cerebroside-3-sulfate, a sphingolipid sulfate ester and a major constituent of the myelin sheet (1). The 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 human ARSA consists of a signal peptide (residues 1-18) and a mature chain (residues 19-507) (3). Recombinant human ARSA corresponds to the mature chain and has sulfatase activity described above.

References

  1. Lukatela, G. et al. (1998) Biochemistry 37:3654.
  2. Parenti, G. et al. (1997) Curr. Opin. Genet. & Dev. 7:386.
  3. Stein, C. et al. (1989) J. Biol. Chem. 264:1252.

Alternate Names

ARSA, Cerebroside 3-sulfatase

Entrez Gene IDs

410 (Human); 11883 (Mouse)

Gene Symbol

ARSA

UniProt

AAH14210

Additional Arylsulfatase A/ARSA Products

Product Documents for Recombinant Human Arylsulfatase A/ARSA Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Arylsulfatase A/ARSA Protein, CF

For research use only

Citations for Recombinant Human Arylsulfatase A/ARSA Protein, CF

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Protocols

View specific protocols for Recombinant Human Arylsulfatase A/ARSA Protein, CF (2485-SU):

Materials
  • Assay Buffer: 50 mM NaOAc, 0.5 M NaCl, pH 4.5
  • Recombinant Human Arylsulfatase A/ARSA (rhARSA) (Catalog # 2485-SU)
  • Substrate: 4-Nitrocatechol Sulfate (PNCS) (Sigma, Catalog # N-7251), 100 mM stock in diH2O
  • Sodium Hydroxide (NaOH), 2.0 M stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhARSA to 20 µg/mL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Mix 75 µL of 20 µg/mL of rhARSA and 75 µL of 2 mM of Substrate. As a Substrate Blank combine 75 µL Assay Buffer with 75 µL Substrate only. Prepare in triplicate.
  4. Incubate at 37 °C for 1 hour.
  5. Stop reaction by adding 150 µL of 0.2 M NaOH to reaction vials.
  6. Load in a clear 96-well plate 200 µL from each reaction vial.
  7. Read at 510 nm (absorbance) 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 4-Nitrocatechol (PNC) (Sigma, Catalog # N15553).

Per Well:
  • rhARSA: 1 µg
  • PNCS: 0.5 mM

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