| Description: | A fluorogenic, succinylated peptide substrate for Sirtuin 5. This substrate should be used in conjunction with SIRT Developer, #50089. Optimal excitation wavelength 350-380 nm, emission wavelength 440-460 nm. |
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| UniProt | Q9NXA8 |
| Synonym(s): | SIRT5 |
| Purity: | >90% by HPLC |
| Supplied As: | DMSO solution (50 |
| Format: | DMSO solution |
| Instructions for use: | ASSAY PROTOCOL: Materials: SIRT Assay Buffer (BPS # 50090) SIRT5 Substrate (# 50126) SIRT5 enzyme (# 50016) or other SIRT5-containing sample NUNC black 96-well plate, low protein binding (VWR #62408-936). SIRT Assay Developer (BPS # 50089) Step 1: Set up 50 μl of Sirt5 reactions containing SIRT assay buffer, 400 ng SIRT5 enzyme, and 20 μM SIRT5 substrate in the 96-well plate. Incubate at 37ºC for 30 min. Step 2: Add 50 μl of SIRT Assay Developer (2x) (BPS #50089) to each well and incubate the plate at room temperature for 15 min. 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 (no enzyme control) is subtracted from all other values. |
| Storage / Stability: | Stable at least 6 months from date of receipt, when stored as directed at -20°C or -80°C |
| Application(s): | Suitable for use with BPS |
| Reference(s): | 1. Madsen, A.S. and Olsen, C.A., J. Med. Chem. 2012, 55:5582-5590. 2. Du, J., et al., Science, 11 November 2011, 334:806-809. |
| Warning(s): | Avoid freeze/thaw cycles. Protect from light. |
| Scientific Category: | Deacetylase(s) |
| Product Type: | Substrates |