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

Catalog # Availability Size / Price Qty
4119-GH-010
SDS-PAGE gel picture of purified Recombinant Human alpha-L-Iduronidase / IDUA Protein
1 Image
Product Details
Citations (2)
FAQs
Reviews (1)

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

Product Specifications

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.
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.
Source
Mouse myeloma cell line, NS0-derived human alpha-L-Iduronidase/IDUA protein
Ala26-Pro653 (Ala26Thr), with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Analysis
Thr26
Predicted Molecular Mass
71 kDa
SDS-PAGE
83 kDa, reducing conditions

Product Datasheets

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.

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.

Assay Procedure

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

Data Image

SDS-PAGE SDS-PAGE of Human alpha-L-Iduronidase / IDUA Protein View Larger

2 μg/lane of Recombinant Human alpha-L-Iduronidase / IDUA Protein (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.

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: 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.
Entrez Gene IDs
3425 (Human); 15932 (Mouse); 360904 (Rat)
Alternate Names
alphaLIduronidase; alpha-L-Iduronidase; IDA; IDUA

Citations for Recombinant Human alpha-L-Iduronidase/IDUA 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. Dose-Dependent Prevention of Metabolic and Neurologic Disease in Murine MPS II by ZFN-Mediated In�Vivo Genome Editing
    Authors: K Laoharawee, RC DeKelver, KM Podetz-Ped, M Rohde, S Sproul, HO Nguyen, T Nguyen, SJ St Martin, L Ou, S Tom, R Radeke, KE Meyer, MC Holmes, CB Whitley, T Wechsler, RS McIvor
    Mol. Ther., 2018;26(4):1127-1136.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Bioassay
  2. Shutdown of ER-associated degradation pathway rescues functions of mutant iduronate 2-sulfatase linked to mucopolysaccharidosis type II
    Authors: Y Osaki, A Saito, S Kanemoto, M Kaneko, K Matsuhisa, R Asada, T Masaki, K Orii, T Fukao, S Tomatsu, K Imaizumi
    Cell Death Dis, 2018;9(8):808.
    Species: Human
    Sample Types: Whole Cells
    Applications: Enzyme Assay

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

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Recombinant Human alpha-L-Iduronidase/IDUA Protein, CF
By Anonymous on 05/23/2016
Application: Enzymatic activity in vitro