Ubiquitin-AMC is a fluorogenic substrate for ubiquitin hydrolases based on the C-terminus derivatization of ubiquitin with 7-amido-4-methylcoumarin (AMC). Upon incubation with a protease recognizing Ubiquitin, such as USP2 or UCHL3, AMC is released and the increase in fluorescence can be measured using Ex380 nm and Em460 nm wavelengths. This protein contains no extraneous tags.

P09936

Ub-AMC

≥95% by HPLC.

Typical enzyme concentrations for UCH-L3 are 10-100 pM and for Isopeptidase-T are 10-100 nM.

Release of AMC fluorescence by UCH enzymes can be monitored using Ex380 nm and Em460 nm wavelengths, respectively.

Soluble in DMSO or aqueous buffers (> 1 mg/ml)

Lyophilized solid

Store in the dark at or below –80°C. Stable as supplied for up to 1 year when stored dessicated at –80°C. Store DMSO solutions at –80°C for up to 1 month. Avoid freeze/thaw cycles of solutions.

Useful as a substrate for deubiquitinating enzymes (DUBs), including ubiquitin C-terminal hydrolases (UCHs) and ubiquitin specific proteases (USPs), as well as investigation of deconjugating enzyme substrate specificity and for screening for modulators of

1. Dang, L.C., et al. Kinetic and mechanistic studies on the hydrolysis of ubiquitin C-terminal 7-amido-4-methylcoumarin by deubiquitinating enzymes. Biochemistry, 37, 1868-1879 (1998).
2. Mason, D.E., et al. Substrate profiling of deubiquitin hydrolases with a positional scanning library and mass spectrometry. Biochemistry, 43, 6535-44 (2004).

Protect from light

MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG

Ubiquitination Substrate

Substrate