Recombinant Human IMP Dehydrogenase 1/IMPDH1 Protein, CF

R&D Systems | Catalog # 8904-DH

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human IMP Dehydrogenase 1/IMPDH1 Protein (8904-DH)
  • 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

NP_899066

Applications

Enzyme Activity
View all IMP Dehydrogenase 1/IMPDH1 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human IMP Dehydrogenase 1/IMPDH1 protein
Glu2-Tyr563, with a N-terminal Met and 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

Met

Predicted Molecular Mass

61 kDa

SDS-PAGE

53-64 kDa, reducing conditions

Activity

Measured by its ability to convert the substrate inosine-5'-phosphate (IMP) to xanthosine-5'-phosphate (XMP).
The specific activity is >100 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

8904-DH
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, EDTA, Glycerol and TCEP.
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: IMP Dehydrogenase 1/IMPDH1

IMPDH1 (inosine monophosphate dehydrogenase) is a ubiquitous cytosolic and nuclear enzyme that plays a central role in guanine nucleotide metabolism. It catalyzes the NAD-dependent conversion of inosine monophosphate (IMP) to hypoxanthine monophosphate (XMP) which is a precursor for the synthesis of GMP, guanosine, and guanine. These compounds are critical for DNA synthesis and cell proliferation (1, 2). IMPDH1 associates into a homotetramer of approximately 55 kDa subunits, and the tetramers can aggregate into perinuclear structures (3, 4). IMPDH1 binds to the nucleotides AMP, ATP, IMP, and XMP (4) as well as to single stranded DNA and RNA (5). It is inhibited by the immunosuppressant drug mycophenolic acid (MPA) (3, 6) which induces reversible IMPDH1 aggregation (7). It is required for vascular endothelial cell proliferation and the terminal differentiation of adipocytes (8, 9). Mutations of IMPDH1 are associated with autosomal dominant retinitis pigmentosa (4, 10). Human IMPDH1 shares 98% amino acid sequence identity with mouse and rat IMPDH1. Alternative splicing generates additional isoforms with internal deletions or variable substitutions at the N-terminus.

References

  1. Lane, A.N. and T.W. Fan (2015) Nucleic Acids Res. 43:2466.
  2. Jackson, R.C. et al. (1975) Nature 256:331.
  3. Carr, S.F. et al. (1993) J. Biol. Chem. 268:27286.
  4. Thomas, E.C. et al. (2012) PLoS One 12:e51096.
  5. McLean, J.E. et al. (2004) Biochem. J. 379:243.
  6. Sintchak, M.D. et al. (1996) Cell 85:921.
  7. Ji, Y. et al. (2006) J. Biol. Chem. 281:206.
  8. Chong, C.R. et al. (2006) J. Med. Chem. 49:2677.
  9. Su, H. et al. (2009) Biochem. Biophys. Res. Commun. 386:351.
  10. Bowne, S.J. et al. (2002) Hum. Mol. Genet. 11:559.

Long Name

Inosine 5'-Monophosphate Dehydrogenase 1

Alternate Names

IMPD1, IMPDH1, LCA11, RP10

Entrez Gene IDs

3614 (Human); 23917 (Mouse); 362329 (Rat)

Gene Symbol

IMPDH1

UniProt

NP_899066

Additional IMP Dehydrogenase 1/IMPDH1 Products

Product Documents for Recombinant Human IMP Dehydrogenase 1/IMPDH1 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 IMP Dehydrogenase 1/IMPDH1 Protein, CF

For research use only

Related Research Areas

Cancer Biomarkers
Redox Enzymes

Citations for Recombinant Human IMP Dehydrogenase 1/IMPDH1 Protein, CF

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Protocols

View specific protocols for Recombinant Human IMP Dehydrogenase 1/IMPDH1 Protein, CF (8904-DH):

Materials
  • Assay Buffer: 50 mM Tris, 300 mM NaCl, pH 8.0
  • Recombinant Human IMP Dehydrogenase 1/IMPDH1 (rhIMPDH1) (Catalog # 8904-DH)
  • Nicotinamide adenine dinucleotide sodium salt ( beta -NAD) (Sigma, Catalog # N6522), 100 mM stock in deionized water
  • Inosine 5'-monophosphate (IMP) (Sigma, Catalog # I4625), 100 mM stock in deionized water
  • UV plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhIMPDH1 to 20 μg/mL in Assay Buffer.
  2. Create Substrate Mixture containing 200 µM IMP and 400 µM beta -NAD in Assay Buffer.
  3. Load 50 μL of 20 μg/mL rhIMPDH1 into a plate, and start the reaction by adding 50 μL of Substrate Mixture. For Substrate Blank, load 50 μL of Assay Buffer and 50 μL of Substrate Mixture.
  4. Read plate at a wavelength of 340 nm (bottom read) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

      *Adjusted for Substrate Blank
      **Using the extinction coefficient 6220 M-1cm-1
      ***Using the path correction 0.320 cm
      Note: the output of many spectrophotometers is in mOD Per Well:
  • rhIMPDH1: 1.0 μg
  • beta -NAD: 200 µM
  • IMP: 100 µM

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