Recombinant Mouse alpha-L-Iduronidase/IDUA Protein, CF

R&D Systems | Catalog # 9348-GH

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Key Product Details

  • R&D Systems NS0-derived Recombinant Mouse alpha-L-Iduronidase/IDUA Protein (9348-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

NS0

Accession Number

P48441

Applications

Enzyme Activity
View all alpha-L-Iduronidase/IDUA Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse alpha-L-Iduronidase/IDUA protein
Glu17-Ser634, 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

Glu17

Predicted Molecular Mass

70 kDa

SDS-PAGE

83-95 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic substrate, 4-Methylumbelliferyl  alpha -L-iduronide.
 The specific activity is >7,500 pmol/min/ug, as measured under the described conditions.

Formulation, Preparation, and Storage

9348-GH
Formulation Supplied as a 0.2 μm filtered solution in Sodium Acetate, NaCl and Glycerol.
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-L-Iduronidase/IDUA

alpha -L-Iduronidase encoded by the IDUA gene is an important enzyme required for the lysosomal degradation of glycosaminoglycans (GAGS). It hydrolyzes the
non-reducing terminal alpha -L-iduronic acid residues in GAGS including dermatan sulfate and heparan sulfate.  Mature mouse IDUA shares 80% aa identity with human IDUA. Mutations in IDUA that result in enzymatic deficiency lead to the autosomal recessive disease mucopolysaccharidosis type I (MPS I) (1). MPS I can be classified as three clinical subtypes; Hurler syndrome, Hurler-Scheie syndrome, and Scheie syndrome with decreasing severity, respectively. MPS I causes progressive cellular, tissue and organ damage, and several clinical studies using enzyme replacement therapy show positive results (2, 3). Recently, the IDUA gene has been linked to osteoporosis (4, 5).

References

  1. Scott, H.S. et al. (1995) Hum. Mutat. 6:288.
  2. Wraith, J.E. (2005) Expert Opin. Pharmacother. 6:489.
  3. Jameson, E. (2016) Cochrane Database Syst. Rev. 4: CD009354.
  4. Kodric, K. et al. (2016) Wien Klin Wochenschr. 128:480.
  5. Niu, T. et al. (2016) J. Bone Miner. Res. 31:358.

Alternate Names

alphaLIduronidase, IDA, IDUA

Entrez Gene IDs

3425 (Human); 15932 (Mouse); 360904 (Rat); 102116889 (Cynomolgus Monkey)

Gene Symbol

IDUA

UniProt

P48441

Additional alpha-L-Iduronidase/IDUA Products

Product Documents for Recombinant Mouse alpha-L-Iduronidase/IDUA Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Mouse alpha-L-Iduronidase/IDUA Protein, CF

For research use only

Citations for Recombinant Mouse alpha-L-Iduronidase/IDUA Protein, CF

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Protocols

View specific protocols for Recombinant Mouse alpha-L-Iduronidase/IDUA Protein, CF (9348-GH):

Materials
  • Assay Buffer: 50 mM NaOAc, 150 mM NaCl, 0.02% Brij-35 (w/v), pH 3.5
  • Developing Buffer: 0.1 M Tris, pH 9.0
  • Recombinant Mouse alpha -L-Iduronidase/IDUA (rmIDUA) (Catalog # 9348-GH)
  • Substrate: 4-methylumberlliferyl-alpha -L-Iduronide (Glycosynth, Catalog # 44076), 20 mM stock in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rmIDUA to 0.2 µg/mL in Assay Buffer. Minimize the number of dilution steps to obtain the best activity results.
  2. Dilute Substrate to 200 µM in Assay Buffer.
  3. Combine equal volumes of 0.2 µg/mL rmIDUA and 200 µM Substrate. Include a Substrate Blank containing Assay Buffer and Substrate.
  4. Incubate for 10 minutes at room temperature.
  5. Dilute mixtures to 0.005 µg/mL rmIDUA in Developing Buffer.
  6. In a plate load 100 µL of diluted mixtures.
  7. Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), respectively in endpoint mode.
  8. Calculate specific activity:

Specific Activity (pmol/min/µg) =

Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)
Incubation time (min) x amount of enzyme (µg)

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

Per Well:
  • rmIDUA: 0.0005 µg
  • Substrate: 5 µM

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