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Encompassing Amino Acids: 193- 741
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 25 mM Tris/HCl, pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, and 0.1 mg/ml BSA, 20 µM BPS HDAC substrate 1 (Catalog #50032), 500 µM NAD+ and Sirtuin 1 (12.5 - 800 ng). Incubation condition: 30 min at 37°C followed by treatment with 50 µl of SIRT developer (Catalog # 50089) for 15 min at RT. Fluorescence intensity is measured at ex360/em460.
Description: Human Sirtuin 1 , GenBank Accession No. NM_012238, a.a. 193-741 with N-terminal GST-tag, MW= 87.2 kDa, expressed in an E. coli expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 16 mM glutathione, 20% glycerol
Genbank: NM_012238
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: Q96EB6
Warnings: Avoid freeze/thaw cycles. Storing diluted enzyme is not recommended. If necessary, use carrier protein (BSA 0.1 - 0.5%).
Biosafety Level: Not applicable (BSL-1)
References: 1. Kolthur-Seetharam, U. et al (2006) Cell Cycle 5, 873-877.
2. Solomon, J.M. et al (2006) Mol. Cell. Biol. 26, 28-38.
Application References
1. Arcand, M., et al. "Development of High-Throughput Assays to Study Methylases, Demethylases and Deacetylases." Posterpresented as part of the BioTechniques Fall 2011 Epigenetics Meeting.
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