Tube 1.

Purified Component	Concentration of stock	Volume	Final concentration in assay
BSA	10 mg/mL	0.5 ul	0.25 mg/mL
MgCl2ATP	50 mM	2 ul	5 mM
Substrate	10 uM	1 uL	500 nM
UBCH10	2.5 uM	2 ul	250 nM
UBE2S	2.5 uM	2 ul	250 nM
APC/C	150 nM	2 ul	15 nM
Cdh1	20 uM	1 ul	1 uM
Buffer	1x	5.5 ul	-
Total volume: 16 ul