Recombinant Human GALNS Protein, CF

Catalog # Availability Size / Price Qty
8269-SU-050
R&D Systems Recombinant Proteins and Enzymes
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Recombinant Human GALNS Protein, CF Summary

Product Specifications

Purity
>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to hydrolyze the substrate 4-Nitrocatechol Sulfate (PNCS). The specific activity is >0.5 pmol/min/μg, as measured under the described conditions.
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human N-Acetylgalactosamine-6-Sulfatase/GALNS protein
Ala27-His522, with an N-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
His & Thr56
Predicted Molecular Mass
56 kDa
SDS-PAGE
53-59 kDa, reducing conditions

Product Datasheets

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8269-SU

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.

8269-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.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Sodium Actetate, 250 mM NaCl,  pH 5.0
  • Recombinant Human N‑Acetylgalactosamine-6-Sulfatase/GALNS (rhGALNS) (Catalog # 8269-SU)
  • Substrate: 4-Nitrocatechol Sulfate (PNCS) (Sigma, Catalog # N-7251), 100 mM stock in deionized water
  • Sodium Hydroxide (NaOH) (Sigma, Catalog # 221465), 2 M stock in deionized water
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhGALNS to 40 µg/mL in Assay Buffer.
  2. Dilute PNCS to 4 mM in Assay Buffer.
  3. Combine 200 µL of 40 µg/mL of rhGALNS and 200 µL of 4 mM of PNCS in microtubes.  Include a Substrate Blank with 200 µL of Assay Buffer and 200 µL of 4 mM of PNCS.
  4. Incubate at 37 °C overnight (16-20 hours).
  5. Load 100 µL from each reaction and Substrate Blank vial into a plate in triplicate, minimally.
  6. Stop reaction by adding 100 µL of 0.2 M NaOH to each well containing reactions and Substrate Blank.
  7. Read plate 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:
  • rhGALNS: 2 µg
  • Substrate: 1 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: N-Acetylgalactosamine-6-Sulfatase/GALNS

GALNS is a lysosomal sulfatase that hydrolyzes the 6-sulfate groups of the N-acetyl-D-galactosamine 6-sulfate units of chondroitin sulfate and of the D-galactose 6-sulfate units of keratan sulfate (1). The conversion to 3-oxoalanine (also known as C-formylglycine, FGly) of a cysteine residue is critical for catalytic activity of the enzyme (2). Deficiencies of this enzyme lead to Morquio A syndrome or mucopolysaccharidosis 4A (MPS4A), a lysosomal storage disorder characterized by intracellular accumulation of keratan sulfate and chondroitin-6-sulfate (3). Key clinical features of this disease include short stature, skeletal dysplasia, dental anomalies, and corneal clouding (4). Current therapeutic method for lysosomal storage diseases is enzyme replacement therapy (5).

References
  1. Tomatsu, S. et al. (1991) Biochem. Biophys. Res. Commun. 181:677.
  2. Dierks, T. et al. (1997) Proc. Natl. Acad. Sci. U. S. A. 94:11963.
  3. Rivera-Colon, Y. et al. (2013) J. Mol. Biol. 423:736.
  4. Fukuda, S. et al. (1992) J. Clin. Invest. 90:1049.
  5. Ohashi, T. (2012) Pediatr. Endocrinol. Rev. 10 Suppl 1:26.
Entrez Gene IDs
2588 (Human); 50917 (Mouse); 292073 (Rat)
Alternate Names
Chondroitinase; Chondroitinsulfatase; EC 3.1.6; EC 3.1.6.4; FLJ17434; FLJ42844; FLJ98217; GA6S; galactosamine (N-acetyl)-6-sulfate sulfatase; Galactose-6-Sulfate Sulfatase; GalNAc6S Sulfatase; GALNAC6S; GALNS; GAS; MPS4A; NAcetylgalactosamine6Sulfatase; N-Acetylgalactosamine-6-Sulfatase; N-acetylgalactosamine-6-sulfate sulfatase

Citations for Recombinant Human GALNS 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.

2 Citations: Showing 1 - 2
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  1. Sex Difference Leads to Differential Gene Expression Patterns and Therapeutic Efficacy in Mucopolysaccharidosis IVA Murine Model Receiving AAV8 Gene Therapy
    Authors: M Piechnik, PC Amendum, K Sawamoto, M Stapleton, S Khan, N Fnu, V Álvarez, AMH Pachon, O Danos, JT Bruder, S Karumuthil, S Tomatsu
    International Journal of Molecular Sciences, 2022;23(20):.
    Species: Human
    Sample Types: Plasma
    Applications: ELISA Capture
  2. Bio-Plex immunoassay measuring the quantity of lysosomal N-acetylgalactosamine-6-sulfatase protein in dried blood spots for the screening of mucopolysaccharidosis IVA in newborn: a pilot study
    Authors: CK Chuang, HY Lin, TJ Wang, SF Huang, SP Lin
    BMJ Open, 2017;7(7):e014410.
    Species: Human
    Sample Types: Plasma
    Applications: Standard

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