Detection of Human Sirtuin 5/SIRT5 by Western Blot.
Western blot shows lysates of human liver tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Sirtuin 5/SIRT5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5914) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Sirtuin 5/SIRT5 at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Sirtuin 5/SIRT5 in Human Liver.
Sirtuin 5/SIRT5 was detected in immersion fixed paraffin-embedded sections of human liver using Sheep Anti-Human Sirtuin 5/SIRT5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5914) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to hepatocyte cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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: Sirtuin 5/SIRT5
Sirtuin 5 (SIRT5; SIR2-like protein 5) is a 28‑35 kDa class III member of the sirtuin protein family. It is found in the mitochondrial matrix and is involved in ammonia detoxification. In particular, it deacetylates carbamoyl phosphate synthetase 1/CPS1, thus activating it and allowing for ammonia condensation with bicarbonate and ATP. This is a crucial step in urea formation during prolonged fasting or the use of a high-protein diet. Human SIRT5 is 310 amino acids (aa) in length. It possesses a sertuin‑type deacetylase domain (aa 41‑309) that contains an NAD binding motif (Thr276:Glu277:Pro292). There are three potential isoform variants that show aa substitutions at the C‑terminus. One contains a 14 aa substitution for aa 286‑310, a second shows a 15 aa substitution for aa 287‑310, and the third shows a 17 aa substitution for aa 207‑310. Full-length human and mouse SIRT5 share 86% aa identity.
Silent Mating Type Information Regulation 5 Homolog
Entrez Gene IDs:
23408 (Human); 68346 (Mouse); 306840 (Rat)
EC 3.5.1; EC 3.5.1.-; FLJ36950; NAD-dependent deacetylase sirtuin-5; silent mating type information regulation 2, S.cerevisiae, homolog 5; SIR2L5; sir2-like 5; SIR2-like protein 5; SIRT5; sirtuin (silent mating type information regulation 2 homolog) 5 (S. cerevisiae); sirtuin (silent mating type information regulation 2, S.cerevisiae, homolog) 5; Sirtuin 5; sirtuin type 5
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