Table 1
(continued)
Khan et al.
(2018 ) [13]
Lin et al.
(2018) [12]
Conlan et al.
(2018) [14]
Zhang et al.
(2019) [16]
a
Mair et al.
(2019) [17]
b
Kim et al.
(2019) [18]
c
Biotin
treatment:
Concentration
and duration
2 mM for 24 h
50
μM for 24 h
75
μM for 24 h
200
μM for 12 h
for TurboID;
200
μM for 24 h
for BioID and
BioID2 at 37
C
2.5–50
μM for
TurboID for 0.5
and/or 3 h
e
;
20–50
μM for
miniTurbo for 1 h
e
50
μM for 3 h
Removal of
excess biotin
Amicon Ultra-4
centrifugal filter
columns (Millipore
Sigma)
Three washes with
0.6 M mannitol
solution
PD10 size exclusion
column
(GE Healthcare)
Zeba Spin
Desalting
column
(ThermoFisher
Scientific)
1 or 2 PD-10 gel
filtration columns
(GE-Healthcare)
Sephadex G-25 in
PD-10 desalting
columns
(GE Healthcare)
Purification of
biotinylated
proteins
30
μL StreptavidinSepharose “high
performance” beads
(GE healthcare)
rotated overnight at
4
C
600
μL Dynabeads
MyOne
Streptavidin C1
(Invitrogen)
incubated
overnight at 4
C
300
μL Dynabeads
MyOne
Streptavidin C1
(ThermoFisher
Scientific) rotated
at 4
C for 3 h
200
μL Dynabeads
MyOne
Streptavidin C1
(Invitrogen)
rotated
overnight at 4
C
200
μL Dynabeads
MyOne Streptavidin
C1 (Invitrogen)
rotated overnight at
4
C
30
μL Dynabead C1
Streptavidin beads
(ThermoFisher
Scientific)
incubated at 4
C
for 3 h
a
Zhang et al. (2019) [16] used 200
μM biotin for 12 h with TurboID in
N. benthamiana for final mass spectrometry analysis
b
Mair et al. (2019) [17] used 50
μM biotin with TurboID in Arabidopsis for final mass spectrometry analysis
c
Kim et al. (2019) [18] used TurboID in Arabidopsis for final mass spectrometry analysis
d
Vacuum infiltration not necessary in Arabidopsis seedlings, except unopened floral buds [17]
e
Protein labeling was observed after 10 min for both TurboID and miniTurbo [17]
428
Madiha Khan et al.