Detection of Human ADP‑Sugar Pyrophosphatase/ NUDT5 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, HepG2 human hepatocellular carcinoma cell line, and human liver tissue. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human ADP‑Sugar Pyrophosphatase/NUDT5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6414) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ADP‑Sugar Pyrophosphatase/NUDT5 at approximately 33 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: ADP-Sugar Pyrophosphatase/NUDT5
ADP‑Sugar Pyrophosphatase (NUDT-5, Nucleoside diphosphate-linked moiety X motif 5; also YSA1H and 8-oxo-dGDP phosphatase) is a member of the Nudix hydrolase family of proteins. Although the predicted MW of NUDT-5 is 24 kDa, it runs anomalously at 32-36 kDa in SDS-Page. NUDT-5 is a widely expressed intracellular homodimer that has at least two functions. First, it possesses pyrophosphatase activity that prevents nonenzymatic ribosylation of proteins, and second, it hydrolyzes 8-oxoGDP (plus 8-OH-dADP), thus removing an oxidized nucleoside from the nucleotide pool and its potential for incorporation into either DNA or RNA. Such incorporation likely results in transcriptional or translational errors. Human NUDT-5 is 219 amino acids (aa) in length (SwissProt #:Q9UKK9). It is contains an N-terminal substrate binding region (aa 28-51), a Nudix hydrolase domain (aa 57-197) and an extended C-terminal a-helix (aa 211-219) NUDT-5 is acetylated on Met1, Lys42, Lys210, and Lys218, and phosphorylated on Tyr74. There are two isoform variants, one that shows a deletion of aa 163-165, and another that contains a 13 aa substitution for aa 164-219. Full-length human and mouse NUDT-5 share 82% aa sequence identity.
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