Sirtuin 2, encoded by the SIRT2 gene, is also known as SIR2 (silent information regulator 2)-like protein 2. It is a nicotinamide adenine dinucleotide (NAD)-dependent histone/protein deacetylase (1, 2). The SIR2 family of enzymes is classified as class III histone deacetylases (HDACs) and has been implicated in many cellular processes that include histone deacetylation, gene silencing, chromosomal stability, and aging (3, 4). Unlike class I and class II HDACs, the enzymatic activity of class III HDACs is NAD dependent and insensitive to HDAC inhibitor trichostatin A (5).
Recombinant Human Sirtuin 2/SIRT2 Protein, CF
R&D Systems | Catalog # 4358-DA
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Key Product Details
- R&D Systems Sf 21 (baculovirus)-derived Recombinant Human Sirtuin 2/SIRT2 Protein (4358-DA)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
Sf 21 (baculovirus)
Accession Number
Applications
Enzyme Activity
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Product Specifications
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived human Sirtuin 2/SIRT2 protein
Met1-Gln389, with a C-terminal 6-His tag
Met1-Gln389, with a C-terminal 6-His tag
Purity
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
No results obtained: Presumed blocked
Predicted Molecular Mass
44 kDa
SDS-PAGE
43-49 kDa, reducing conditions
Activity
Measured by its ability to remove the acetyl group from a fluorogenic peptide substrate Ac-RGK(Ac)-AMC (Catalog # ES016) in a coupled assay.
The specific activity is >3 pmol/min/µg, as measured under the described conditions.
The specific activity is >3 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
4358-DA
| Formulation | Supplied as a 0.2 μm filtered solution in Tris 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.
|
Background: Sirtuin 2/SIRT2
References
- Frye, R.A. (1999) Biochem. Biophys. Res. Commun. 260:273.
- North, B.J. et al. (2003) Mol. Cell. 11:437.
- North, B.J. and Verdin, E. (2004) Genome Biol. 5:224.
- Marmorstein, R. (2004) Biochem. Soc. Trans. 32:904.
- Gray, S.G. and Ekstrom, T.J. (2001) Exp. Cell Res. 262:75.
Long Name
Silent Mating Type Information Regulation 2 Homolog
Alternate Names
SIR2L2, SIRT2
Gene Symbol
SIRT2
UniProt
Additional Sirtuin 2/SIRT2 Products
Product Documents for Recombinant Human Sirtuin 2/SIRT2 Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human Sirtuin 2/SIRT2 Protein, CF
For research use only
Related Research Areas
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Protocols
View specific protocols for Recombinant Human Sirtuin 2/SIRT2 Protein, CF (4358-DA):
Materials
- Assay Buffer: 25 mM Tris, 150 mM NaCl, 1 mM DTT, pH 8.0
- Stop Solution: 8 ng/µL Recombinant Human Active Trypsin 3/PRSS3 (Catalog # 3714-SE), 4 mM Nicotinamide (Sigma, Catalog # 72340), 50 mM Tris, 100 mM NaCl, 30% (v/v) isopropanol, pH 8.0
- Recombinant Human Sirtuin 2/SIRT2 (rhSIRT2) (Catalog # 4358-DA)
- beta -Nicotinamide adenine dinucleotide hydrate ( beta -NAD) (Sigma, Catalog # N6522), 100 mM stock in diH2O
- Substrate: Ac-Arg-Gly-Lys(Ac)-AMC (Catalog # ES016), 20 mM stock in DMSO
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rhSIRT2 to 0.04 µg/µL in Assay Buffer.
- Dilute Substrate to 500 µM in Assay Buffer containing 2 mM beta -NAD.
- Mix 25 µLof 0.04 µg/µL rhSIRT2 and 25 µL of Substrate in plate. Add 25 µL of 0.04 µg/µL rhSIRT2 alone for the control.
- Cover with parafilm or a plate sealer and incubate at 37 °C for 30 minutes.
- Add 50 µL of Stop Solution to all wells. Mix. For control well(s), add 25 µL of Substrate after addition of Stop Solution.
- Seal tightly and incubate at room temperature for 15 minutes.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively in endpoint mode.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU) |
| Incubation time (min) x amount of enzyme (µg) |
*Adjusted for Control
**Derived using calibration standard 7-amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).
Per Well:
- rhSIRT2: 1 µg
- Substrate: 125 µM
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