Recombinant Human Arylsulfatase A/ARSA Protein, CF Summary
Arg19-Ala507, with a C-terminal 10-His tag
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: 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
- Dilute rhARSA to 20 µg/mL in Assay Buffer.
- Dilute Substrate to 2 mM in Assay Buffer.
- 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.
- Incubate at 37 °C for 1 hour.
- Stop reaction by adding 150 µL of 0.2 M NaOH to reaction vials.
- Load in 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).Per Well:
- rhARSA: 1 µg
- PNCS: 0.5 mM
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.
- Lukatela, G. et al. (1998) Biochemistry 37:3654.
- Parenti, G. et al. (1997) Curr. Opin. Genet. & Dev. 7:386.
- Stein, C. et al. (1989) J. Biol. Chem. 264:1252.
Citation for Recombinant Human Arylsulfatase A/ARSA 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
Substrate reduction therapy for Krabbe disease and metachromatic leukodystrophy using a novel ceramide galactosyltransferase inhibitor
Authors: MC Babcock, CR Mikulka, B Wang, S Chandriani, S Chandra, Y Xu, K Webster, Y Feng, HR Nelvagal, A Giaramita, BK Yip, M Lo, X Jiang, Q Chao, JC Woloszynek, Y Shen, S Bhagwat, MS Sands, BE Crawford
Scientific Reports, 2021;11(1):14486.
Sample Types: Recombinant Protein
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