Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein, CF

R&D Systems | Catalog # 8349-DH

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein (8349-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

P12268

Applications

Enzyme Activity
View all IMP Dehydrogenase 2/IMPDH2 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human IMP Dehydrogenase 2/IMPDH2 protein
Met1-Phe514, with a C-terminal 10-His tag

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met1 predicted

Predicted Molecular Mass

57 kDa

SDS-PAGE

52-61 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 >30 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein, CF

Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein Enzyme Activity

Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein Enzyme Activity

Recombinant Human IMPDH2 (Catalog # 8349-DH) is measured by its ability to convert the substrate inosine-5'-phosphate (IMP) to xanthosine-5'-phosphate (XMP). The activity (orange) is approximately 3.5-fold greater than the competitor's IMPDH2 (green).
Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein SDS-PAGE

Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein SDS-PAGE

1 μg/lane of Recombinant Human Inosine 5'-Monophosphate Dehydrogenase 2/IMPDH2 was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silverstaining, showing a band at 57 kDa.

Formulation, Preparation, and Storage

8349-DH
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT and Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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 2/IMPDH2

IMPDH2 (inosine monophosphate dehydrogenase) is one of two cytosolic and nuclear enzyme isoforms that play a central role in guanine nucleotide metabolism. The isoforms are 84% identical but distinctly regulated. While IMPDH1 is generally constitutively expressed, IMPDH2 is inducible during proliferation and transformation (1, 2). Both isoforms form a homotetramer of approximately 55 kDa monomers containing a catalytic barrel domain and a subdomain with two cystathione beta-synthase domains which mediate RNA and DNA binding (1, 3). Both enzymes catalyze 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 (4, 5) which explains the importance of IMPDH in cancer and viral infection (6-9). Although IMPDH1 and IMPDH2 are known to be inhibited by the immunosuppressant drug mycophenolic acid (MPA), much research has targeted discovery of additional and selective inhibitors for IMPDH isoforms given they are targets for several major therapeutic areas (1, 9-13). Human IMPDH2 shares 99% amino acid sequence identity with mouse IMPDH2.

References

  1. Hedstrom, L. (2009) Chem. Rev. 109:2903.
  2. Thomas, E.C. et al. (2012) PLoS One 12:e51096.
  3. McLean, J.E. et al. (2004) Biochem. J. 379:243.
  4. Lane, A.N. and T.W. Fan (2015) Nucleic Acids Res. 43:2466.
  5. Jackson, R.C. et al. (1975) Nature 256:331.
  6. Zho, J. et al. (2015) Med. Oncol. 32:373.
  7. Xu, Y. et al. (2017) Sci. Rep. 7:745.
  8. Nair, V. (2007) Antivir. Chem. Chemother. 18:245.
  9. Trapero, A. et al. (2018) J. Med. Chem. 61:2806.
  10. Barnes, B.J. et al. (2001) Int. J. Cancer 94:275.
  11. Cholewinksi, G. et al. (2015) J. Enzyme Inhib. Med. Chem. 30:550.
  12. Liao, L.X. et al. (2017) Proc. Natl. Acad. Sci. USA 114:E5986.
  13. Cuny, G.D. et al. (2017) Expert Opin. Ther. Pat. 27:677.

Long Name

Inosine 5'-Monophosphate Dehydrogenase 2

Alternate Names

IMDH2, IMP Oxireductase 2, IMPD2, IMPDH2

Entrez Gene IDs

3615 (Human); 23918 (Mouse); 301005 (Rat)

Gene Symbol

IMPDH2

UniProt

P12268

Additional IMP Dehydrogenase 2/IMPDH2 Products

Product Documents for Recombinant Human IMP Dehydrogenase 2/IMPDH2 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 2/IMPDH2 Protein, CF

For research use only

Related Research Areas

Redox Enzymes

Citations for Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein, CF

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Protocols

View specific protocols for Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein, CF (8349-DH):

Materials
  • Assay Buffer: 50 mM Tris, 300 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 8.0
  • Recombinant Human Inosine 5'-Monophosphate Dehydrogenase 2/IMPDH2 (rhIMPDH2) (Catalog # 8349-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 rhIMPDH2 to 20 μg/mL in Assay Buffer.
  2. Create Substrate Mixture containing 500 μM IMP and 1 mM beta -NAD in Assay Buffer.
  3. Load 50 μL of 20 μg/mL rhIMPDH-2 into a plate, and start the reaction by adding 50 μL of Substrate Mixture. For Substrate Blanks, load 50 μL of Assay Buffer and 50 μL of Substrate Mixture.
  4. Read plate at a wavelength of 339 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:
  • rhIMPDH-2: 1.0 μg
  • beta -NAD: 500 μM
  • IMP: 250 μM

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