glutathione
s-transferase
gst
bcl-gl
recombinant
protein
expressed
escherichia
coli
strain
bl21
codon-plus
ril
competent
cells
stratagene
purification
recombinant
proteins
performed
using
glutathione
sepharose
4b
beads
ge
healthcare
under
nondenaturing
conditions
according
supplier
instructions
confirmation
direct
binding
bcl-gl
melk
removed
gst
from
gst-fused
bcl-gl
protein
using
prescission
protease
ge
healthcare
according
supplier
instructionssoluble
fractions
filtered
syringe
filters
applied
ni-nta
affinity
column
pre-equilibrated
tris-hcl
ph
nacl
β-mercaptoethanol
target
protein
complexes
subunit
gst-tag
sv40
st
his-tag
eluted
elution
buffer
tris-hcl
ph
nacl
imidazole
β-mercaptoethanol
dialyzed
overnight
tris-hcl
ph
nacl
dtt
dialyzed
protein
applied
gst
affinity
column
remove
free
sv40
st
on-column
cleavage
tev
protease
performed
overnight
flow-through
fraction
gst
column
reapplied
ni-nta
column
remove
cleaved
his-tag
tev
protease