Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein, CF

Catalog #: 8349-DH
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
8349-DH-050

Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein Enzyme Activity

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All IMP Dehydrogenase 2/IMPDH2 Proteins and Enzymes

Product Details

Citations (1)

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human IMP Dehydrogenase 2/IMPDH2 Protein, CF Summary

Product Specifications

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.

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.

Source

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

Accession #

P12268

N-terminal Sequence
Analysis

Met1 predicted

Predicted Molecular Mass

57 kDa

SDS-PAGE

52-61 kDa, reducing conditions

Product Datasheets

8349-DH

Product Datasheet

COA

SDS

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.

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.

Assay Procedure

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

Dilute rhIMPDH2 to 20 μg/mL in Assay Buffer.
Create Substrate Mixture containing 500 μM IMP and 1 mM beta -NAD in Assay Buffer.
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.
Read plate at a wavelength of 339 nm (bottom read) in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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

Scientific Data

Enzyme Activity

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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).

SDS-PAGE

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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.

Reconstitution Calculator

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

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

Long Name

Inosine 5'-Monophosphate Dehydrogenase 2

Entrez Gene IDs

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

Alternate Names

EC 1.1.1.205; IMDH2; IMP (inosine 5'-monophosphate) dehydrogenase 2; IMP (inosine monophosphate) dehydrogenase 2; IMP Dehydrogenase 2; IMP Oxireductase 2; IMPD 2; IMPD2; IMPD2IMP oxireductase 2; IMPDH 2; IMPDH2; IMPDH-II; inosine 5' phosphate dehydrogenase 2; inosine monophosphate dehydrogenase type II; inosine-5'-monophosphate dehydrogenase 2

Related Research Areas

Citation for Recombinant Human IMP Dehydrogenase 2/IMPDH2 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.

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