Recombinant Human GALNS Protein, CF

R&D Systems | Catalog # 8269-SU

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human GALNS Protein (8269-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

Sf 21 (baculovirus)

Accession Number

P34059

Applications

Enzyme Activity
View all N-Acetylgalactosamine-6-Sulfatase/GALNS Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human N-Acetylgalactosamine-6-Sulfatase/GALNS protein
Ala27-His522, with an N-terminal 6-His tag

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.

N-terminal Sequence Analysis

His & Thr56

Predicted Molecular Mass

56 kDa

SDS-PAGE

53-59 kDa, reducing conditions

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.

Formulation, Preparation, and Storage

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.

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.

Alternate Names

Chondroitinase, Chondroitinsulfatase, GA6S, Galactose-6-Sulfate Sulfatase, GalNAc6S Sulfatase, GALNS, GAS, MPS4A, NAcetylgalactosamine6Sulfatase

Entrez Gene IDs

2588 (Human); 50917 (Mouse); 292073 (Rat)

Gene Symbol

GALNS

UniProt

P34059

Additional N-Acetylgalactosamine-6-Sulfatase/GALNS Products

Product Documents for Recombinant Human GALNS Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human GALNS Protein, CF

For research use only

Citations for Recombinant Human GALNS Protein, CF

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Protocols

View specific protocols for Recombinant Human GALNS Protein, CF (8269-SU):

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

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