| #REF | SIZE | PRICE | |
|---|---|---|---|
| #E-Nov6-50 | 50 mUnits | € 305.00 | Inquiry |
| #E-Nov6-100 | 150 mUnits | € 650.00 | Inquiry |
Pricing updated on July 23, 2024.
Kit is provided in stable lyophilized form and shipped without dry ice
You can ask us for a quotation here or write at contact@novocib.com
NOVOCIB'S human cytosolic IMP/GMP specific 5'-nucleotidase/phosphotransferase II (cN-II) is a pure and active protein of 65kDa cloned by RT-PCR amplification of mRNA extracted from human hepatoma cells and expressed in E.coli. The sequence of the cloned NT5C2 gene (GenBank accession number P49902) was confirmed by DNA sequencing (100% identity).
Cytosolic 5'-nucleotidase II is one of the seven known mammalian nucleotidases(1) that specifically catalyzes the dephosphorylation of 6-hydroxypurine nucleoside 5'-monophosphates (IMP, dIMP, GMP, dGMP) and regulates cellular pool of IMP and GMP(2,3). The enzyme also acts as a phosphotransferase catalyzing the transfer of a phosphate from nucleoside monophosphate to a nucleoside acceptor ' preferentially inosine and deoxyinosine. Unlike the other 5'-nucleotidases, cN-II is allosterically regulated by adenine/guanine nucleotides and 2,3-biphosphoglycerate(4).
In addition, cytosolic 5'-nucleotidase II phosphorylates anti-viral and anti-tumour nucleoside analogues such as 2'3'-dideoxyinosine, carbovir(5), acyclovir(6) and ribavirin(7).
Synonyms: cytosolic 5'-nucleotidase/phosphotransferase, High Km 5'-nucleotidase (hkm-NT), cytosolic purine 5µ-nucleotidase (purine 5'-NT), IMP/GMP-specific 5µ-nucleotidase
Download Human cytosolic 5'-nucleotidase II (User manual)
Assay condition: Phosphotransferase activity of human recombinant cN-II was followed at 37°C, in 100mM Tris-HCl pH 9; 100mM KCl, 20mM MgCl2, 0.5mM KH2PO4, 5mM DTT, 5mM NAD, 50mU/mL IMPDH (NovoCIB). 5mM GMP was used as phosphate donor and 5mM inosine as - phosphate acceptor. The formation of IMP from inosine was followed at 340nm through coupling to recombinant IMP-dehydrogenase (IMPDH, NovoCIB). In the presence of NAD, IMPDH catalyzes irreversible conversion of IMP to XMP with concomitant reduction of NAD to NADH2 (specific absorbance at 340nm).
Assay condition: 5'-nucleotidase activity of cN-II is followed in an irreversible spectrophotometric assay using coupled purine nucleoside phosphorylase - xanthine dehydrogenase system (2,5 mU/ml each). Assays are carried out at 37'C, at 50mM Tris-HCl pH7,6; 100mM KCl, 20mM MgCl2, 500'M KH2PO4, 5mM DTT, 119 nM cN-II and various concentrations of IMP. Reaction is followed at 295 nm. IMP is purchased from MP Biochemicals. PNP and XDH are produced and purified by NovoCIB.
