A fluorogenic, myristoylated lysine peptide substrate for Sirtuin 6. This substrate should be used in conjunction with SIRT Developer, #50089. Optimal excitation wavelength 350−380 nm, emission wavelength 440−460 nm.

SIRT6

>90% by HPLC

DMSO solution

5 mM

DMSO solution

ASSAY PROTOCOL:

Materials: SIRT Assay Buffer (BPS Bioscience, # 50090) SIRT6 Substrate (BPS Bioscience, #50866) SIRT6 enzyme (BPS Bioscience, #50017) or other SIRT6-containing sample NUNC black 96-well plate, low protein binding (VWR #62408-936). SirtuinDeveloper (2X) (BPS Bioscience, #50089)

Step 1: Set up 50 µl of SIRT6 reactions containing SIRT assay buffer, 1 μg SIRT6 enzyme, and 5 µM SIRT6 substrate in the 96-well plate. Include a no-enzyme control reaction (“Blank”) to determine background fluorescence. Incubate at 37ºC for 30 minutes.

Step 2: Add 50 µl of SIRT Assay Developer (2x) (BPS #50089) and incubate the plate at room temperature for 15 minutes.

Step 3: Read sample in a microtiter-plate reading fluorimeter capable of excitation at a wavelength in the range 350-380 nm and detection of emitted light in the range 440-460 nm. “Blank” value is subtracted from all other values.

Stable at least 6 months from date of receipt, when stored as directed at -20°C or -80°C

Suitable for use with BPS’s Fluorogenic SIRT6 Assay Kit, #50022, and to study enzyme kinetics and screen small molecule inhibitors of Sirtuin 6 for drug discovery and HTS applications.

1. Hu, J. et al., 2013. Org Biomol Chem, 11(32):5213-5216.

2. Rahnasto-Rilla, M., et al., 2016. Methods Mol Biol, 1436:259-269.

Avoid freeze/thaw cycles. Protect from light.

Deacetylase

Substrate