Table 1. DUBs used in UbiCRest.

Linkage type	DUB	Useful final [conc] (1x)	Comments on the DUB	Ref.
All eight linkages including proximal ubiquitin	USP21 or USP2	1-5 µM (USP21)	Positive control.	28
All linkages excluding Met1 and most proximal ubiquitin	CCHFV viral OTU (vOTU)	0.5-3 µM	Positive control, does not cleave Met1 linkages.	38
Lys6	OTUD3	1-20 µM	Cleaves Lys11 chains equally well. Targets other isopeptide linkages at high concentrations (Lys63 better (>) than the others, not Met1).	27,31
Lys11	Cezanne	0.1-2 µM	Very active, non-specific at very high concentrations (Lys63 > Lys48 > others, not Met1).	27,39
Lys27	OTUD2	1-20 µM	Also cleaves Lys11, Lys29, Lys33, prefers longer Lys11 chains. Non-specific at high concentrations (Lys48 > others, not Met1).	27
Lys29	TRABID	0.5-10 µM	Cleaves Lys 29 and Lys33 equally well, and Lys63 with lower activity. Might target other linkages at higher concentrations (not Met1). Low yields from bacterial expression.	40
Lys33
Lys48	OTUB1	1-20 µM	Highly Lys48-specific. Not very active. Can be used at high concentrations.	27,41
Lys63	OTUD1	0.1-2 µM	Very active, non-specific at high concentrations (Lys48 > others, not Met1).	27
Lys63	AMSH / AMSH-LP	1-20 μM	Highly Lys63-specific. Not very active against long chains. Can be used at high concentrations.	5,23
Met1/linear	OTULIN	Any (10 nM – 10 μM)	Very active and specific, can be used at high concentrations.	32