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

R&D Systems | Catalog # 4119-GH

NS0-expressed, analyzed by SEC-MALS
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Key Product Details

  • R&D Systems NS0-derived Recombinant Human alpha-L-Iduronidase/IDUA Protein (4119-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

AAA81589

Applications

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

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human alpha-L-Iduronidase/IDUA protein
Ala26-Pro653 (Ala26Thr), with a C-terminal 10-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

Thr26

Predicted Molecular Mass

71 kDa

SDS-PAGE

83 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/µg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 4119-GH in the following applications:

Scientific Data Images for Recombinant Human alpha-L-Iduronidase/IDUA Protein, CF

Recombinant Human alpha ‑L‑Iduronidase/IDUA SEC-MALS.

Recombinant Human alpha ‑L‑Iduronidase/IDUA (Catalog # 4119-GH) has a molecular weight (MW) of 99-110 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

Recombinant Human alpha-L-Iduronidase/IDUA Protein Activity Assay Diagram.

Recombinant Human alpha-L-Iduronidase/IDUA Protein, CF (Catalog # 4119-GH) hydrolyzes iduronic acid from dermatan and heparan sulfates.

Recombinant Human alpha-L-Iduronidase/IDUA Protein Activity Assay.

Recombinant Human alpha-L-Iduronidase/IDUA Protein, CF (Catalog # 4119-GH) is measured by its ability to cleave a fluorogenic substrate, 4-Methylumbelliferyl alpha -L-iduronide.
SDS-PAGE gel picture of purified Recombinant Human alpha-L-Iduronidase / IDUA Protein

SDS-PAGE of Human alpha-L-Iduronidase / IDUA Protein.

2 μg/lane of Recombinant Human alpha-L-Iduronidase / IDUA Protein (Catalog # 4119-GH) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band at ~83 kDa under reducing conditions.

Formulation, Preparation, and Storage

4119-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. Mutations in IDUA that result in enzymatic deficiency lead to the autosomal recessive disease mucopolysaccharidosis type I (MPS I) (1). MPS I causes progressive cellular, tissue and organ damage, and several clinical studies using enzyme replacement therapy have shown promising benefits (2).

References

  1. Scott, H.S. et al. (1995) Hum. Mutat. 6:288.
  2. Wraith, J.E. (2005) Expert Opin. Pharmacother. 6:489.

Alternate Names

alphaLIduronidase, IDA, IDUA

Entrez Gene IDs

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

Gene Symbol

IDUA

UniProt

AAA81589

Additional alpha-L-Iduronidase/IDUA Products

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

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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

For research use only

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

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Protocols

View specific protocols for Recombinant Human alpha-L-Iduronidase/IDUA Protein, CF (4119-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 Human alpha ‑L‑Iduronidase/IDUA (rhIDUA) (Catalog # 4119-GH)
  • Substrate: 4-methylumberlliferyl-alpha -L-iduronide (Glycosynth, Catalog # 44076), 20 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 rhIDUA 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 rhIDUA 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 in Developing Buffer.
  6. Load 100 µL of the diluted mixtures into a plate.
  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:
  • rhIDUA: 0.0005 µg
  • Substrate: 5 µM

FAQs

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