Recombinant Human IMP Dehydrogenase 1/IMPDH1 Protein, CF

• Reactivity: Hu
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant Human IMP Dehydrogenase 1/IMPDH1 Protein, CF Summary

• Details of Functionality: 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.
• Source: Spodoptera frugiperda, Sf 21 (baculovirus)-derived human IMP Dehydrogenase 1/IMPDH1 protein
  Glu2-Tyr563, with a N-terminal Met and 6-His tag
• Accession #: NP_899066
• N-terminal Sequence: Met
• Protein/Peptide Type: Recombinant Enzymes
• Gene: IMPDH1
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 61 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 53-64 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris, NaCl, EDTA, Glycerol and TCEP.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Assay Procedure:
  • 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 10^12 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human IMP Dehydrogenase 1/IMPDH1 Protein, CF

• DKFZp781N0678
• EC 1.1.1.205
• IMP (inosine 5'-monophosphate) dehydrogenase 1
• IMP (inosine monophosphate) dehydrogenase 1
• IMP Dehydrogenase 1
• IMPD 1
• IMPD
• IMPD1
• IMPDH 1
• IMPDH1
• IMPDH-I
• inosine-5'-monophosphate dehydrogenase 1
• LCA11
• LCA11RP10
• retinitis pigmentosa 10 (autosomal dominant)
• RP10
• sWSS2608

Background

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.

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.

Publications for IMPDH1 (8904-DH)(1)

Publication using 8904-DH

• Lee, TJ;Sasaki, Y;Ruzycki, PA;Ban, N;Lin, JB;Wu, HT;Santeford, A;Apte, RS; Catalytic isoforms of AMP-activated protein kinase differentially regulate IMPDH activity and photoreceptor neuron function JCI insight 2024-01-16 [PMID: 38227383] (Bioassay, N/A)

FAQs for IMPDH1 (8904-DH). (Showing 1 - 1 of 1 FAQ).

1. Do you have a kit for measuring IMPDH activity?
   • We currently do not have any kits to measure IMPDH activity, however we have a number of IMPDH antibody pairs that can be used for WB/IP and ELISA.

Bioinformatics

• Gene Symbol: IMPDH1
• Uniprot:
  • NP_899066()