Recombinant Human alpha-Galactosidase A/GLA Protein, CF

R&D Systems | Catalog # 6146-GH

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human alpha-Galactosidase A/GLA Protein (6146-GH)
  • 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

CHO

Accession Number

P06280

Applications

Enzyme Activity
View all alpha-Galactosidase A/GLA Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human alpha-Galactosidase A/GLA protein
Met1-Leu429, with a C-terminal 6-His tag

Purity

>95%, 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

Leu32

Predicted Molecular Mass

46 kDa

SDS-PAGE

45-55 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze 4-methylumbelliferyl-alpha -D-galactopyranoside.
The specific activity is >1,100 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 2 reviews rated 5 using 6146-GH in the following applications:

Scientific Data Images for Recombinant Human alpha-Galactosidase A/GLA Protein, CF

Recombinant Human alpha-Galactosidase A/GLA Protein Enzyme Activity Diagram.

Recombinant Human alpha-Galactosidase A/GLA Protein, CF (Catalog # 6146-GH) hydrolyses the terminal alpha -galactosyl moieties from glycolipids and glycoproteins. The common ceramide trihexoside substrate is displayed in the diagram.

Recombinant Human alpha-Galactosidase A/GLA Protein Enzyme Activity.

Recombinant Human alpha-Galactosidase A/GLA Protein, CF (Catalog # 6146-GH) is measured by its ability to hydrolyze 4-methylumbelliferyl-alpha -D-galactopyranoside.

Recombinant Human alpha ‑Galactosidase A/GLA Protein SDS-PAGE.

2 μg/lane of Recombinant Human alpha ‑Galactosidase A/GLA Protein (Catalog # 6146-GH) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 45-55 kDa.

Formulation, Preparation, and Storage

6146-GH
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: alpha-Galactosidase A/GLA

Human alpha -Galactosidase A is a homodimeric glycoprotein that can release terminal alpha -galactosyl moieties from glycolipids and glycoproteins and catalyze the hydrolysis of melibiose into galactose and glucose (1). It is a lysosomal enzyme and is responsible for degradation of glycolipid globotriaosylceramide (Gb3) (Gal alpha 1‑4Gal beta 1‑4Glc beta ‑ceramide). Mutations in this gene cause Fabry disease, an X-linked hereditary lysosomal storage disease with the accumulation of Gb3 in the walls of small blood vessels, nerves, dorsal root ganglia, renal glomerular and tubular epithelial cells, and cardiomyocytes (2, 3). Inability to prevent the glycosphingolipid deposition can cause hypertension, strokes, heart attack and progressive renal failure (4). Current treatment for Fabry disease is enzyme replacement therapy using intravenously delivered recombinant alpha -Galactosidase A (5, 6).

References

  1. Ioannou, Y.A. et al. (1998) Biochem. J. 332:789.
  2. Koide, T. et al. (1990) FEBS Lett. 259:353.
  3. Ioannou Y.A, et al. (1992) J. Cell Biol. 119:1137.
  4. Germain, D.P. (2002) Expert. Opin. Investig. Drugs. 11:1467.
  5. Barngrover, D. (2003) J. Biotechnol. 95:280.
  6. Mignani, R. and Cagnoli, L. (2004) J. Nephrol. 17:354.

Alternate Names

Agalsidase alpha, GALA, Melibiase

Entrez Gene IDs

2717 (Human)

Gene Symbol

GLA

UniProt

P06280

Additional alpha-Galactosidase A/GLA Products

Product Documents for Recombinant Human alpha-Galactosidase A/GLA Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human alpha-Galactosidase A/GLA Protein, CF

For research use only

Citations for Recombinant Human alpha-Galactosidase A/GLA Protein, CF

Customer Reviews for Recombinant Human alpha-Galactosidase A/GLA Protein, CF (2)

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    Application: Enzymatic activity in vitro
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Protocols

View specific protocols for Recombinant Human alpha-Galactosidase A/GLA Protein, CF (6146-GH):

Materials
  • Assay Buffer: 50 mM Sodium Citrate, 50 mM NaCl, pH 4.0
  • Recombinant Human alpha ‑Galactosidase A/GLA (rhGLA) (Catalog # 6146-GH)
  • Substrate: 4-methylumbelliferyl-alpha -D-galactopyranoside (Sigma, Catalog # M7633), 6.7 mM Stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhGLA to 4.0 ng/µL in Assay Buffer.
  2. Dilute Substrate to 800 µM in Assay Buffer.
  3. Load into plate 50 µL of 4.0 ng/µL rhGLA, and start the reaction by adding 50 µL of 800 µM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate.
  4. Read at excitation and emission wavelengths of 365 nm and 445 nm, respectively, in kinetic mode for 5 minutes.
  5. Calculate the specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 4-Methylumbelliferone (Sigma, Catalog # M1381).

Per Well:
  • rhGLA: 0.200 µg
  • Substrate: 400 µM

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