Detection of Human Sirtuin 3/SIRT3 by Western Blot.
Western blot shows lysates of human liver tissue, human heart ventricle tissue, and human kidney tissue. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human Sirtuin 3/SIRT3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7488) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Sirtuin 3/SIRT3 at approximately 28 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: Sirtuin 3/SIRT3
SIRT3 (SIR2-like protein 3) is a 28-30 kDa, NAD+-dependent class III member of the sirtuin protein family. It is widely expressed, and found apparently in both the nucleus, where it deacetylates histones, and in the mitochondrial matrix, where it is involved in the regulation of both oxidative stress and the physiology associated with fasting or caloric restriction. Relative to the mitochondria, SIRT3 deacetylates multiple enzyme substrates, including SOD2, IDH2, LCAD, OTC and HMGCS2, an action that results in enzyme activation. Human SIRT3 is 399 amino acids (aa) in length. It contains a mitochondrial targeting sequence within the first 100 aa, followed by a sertuin-type deacetylase domain (aa 126-382) that contains an NAD binding segment (aa 145-165). Although uncertainty exists relative to its intracellular location, it would appear that SIRT3 is initially synthesized as a 42-44 kDa "precursor" in the cytosol. From here, it can translocate into mitochondria where it is cleaved between Arg99Arg100 to generate a 28-30 kDa short form. There is one potential alternative start site at Met143. Over aa 101-399, human and mouse SIRT3 share 84% aa sequence identity.
Silent Mating Type Information Regulation 3 Homolog
Entrez Gene IDs:
23410 (Human); 64384 (Mouse); 293615 (Rat)
EC 3.5.1; EC 3.5.1.-; hSIRT3; mitochondrial nicotinamide adenine dinucleotide-dependent deacetylase; NAD-dependent deacetylase sirtuin-3, mitochondrial; SIR2L3; SIR2L3silent mating type information regulation 2, S.cerevisiae, homolog 3; sir2-like 3; SIR2-like protein 3; SIRT3; sirtuin (silent mating type information regulation 2 homolog) 3 (S. cerevisiae); sirtuin (silent mating type information regulation 2, S.cerevisiae, homolog) 3; Sirtuin 3; sirtuin type 3
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