Table 1. The reaction conditions of different SIRT6 in vitro assays.
| Activation | Peptide substrate | NAD (mM) |
SIRT6 | Reaction volume (μL) |
Inc. time (min) |
Ex-/Em-wl |
|---|---|---|---|---|---|---|
| HPLC |
QTARK(Ac)STGG: 70 μM [6] |
0.5 | 4 μM | 60 | NA | |
|
ARTKQTARK(Ac)STGGK APRKQLA 40 μM [13] |
0.5 | 0.05 μg/μL | 60 | 30 | NA | |
|
| ||||||
| HPLC |
QTARK(Ac)STGGAc: 300 μM[14] |
2 | 4 μM | 40 | 360 | – |
|
KQTARK(Ac)STGGWW: 50 μM [5] |
1 | 10 μM | 30 | 60 | – | |
|
KQTARK(Myr)STGGWW: 50 μM [5] |
0.5 | 1 μM | 30 | 15 | – | |
|
EALPK(Myr)TGGPQWW: 50 μM [5] |
0.5 | 1 μM | 30 | 15 | – | |
|
ARTKQTARK(Ac)STGGK APRKQLA 150 μM [13] |
0.5 | 0.05 μg/μL | 60 | 60 | NA | |
|
| ||||||
|
Frontal
chrom. |
||||||
|
| ||||||
|
Magnetic
beads |
ARTKQTARK(Ac)STGGK APRKQLA 43 μM [11] |
0.2 | – | 50 | 240 | – |
|
| ||||||
|
RYQK(Ac)-AMCa: 320 μM [9] |
3 | 0.09 μg/μL | 50 | 90 | 370/430 nm | |
|
| ||||||
| Fluorogenic |
EALPK(Myr)-AMCa: 10 μM[8] |
1 | 1 μM | 60 | 120 | 360/460 nm |
|
| ||||||
|
RHKK(Ac)-AMCa, b 400 μM |
3 | 0.09 μg/μL | 50 | 45–90 | 355–370/ 430–460 nm |
|
|
| ||||||
| FRET | (DABCYL)- ISGASE(Myr)DIVHSE- (EDANS)G: 10 μM [10] |
1 | 0.5 μM | 60 | 60 | 336/490 |
a
7-amino-4-methylcoumarin
b
Cayman commercial kit