|
RELIC Peptides
Bioaffinity screening Methods:
Random 12 amino acid combinatorial libraries (Ph.D.12TM) and random
constrained 7 amino acid combinatorial libraries (Ph.D.-C7CTM) were
obtained from New England Biolabs (http://www.neb.com/neb/frame_cat.html).
Immobilized tagged versions of the listed ligands (see each ligand for details)
are used for affinity selection using the recommended number of phage particles
(approximately 1011 pfu on a ligand-coated 60 mm plate) as indicated
for each ligand. Phage particles are propagated in E. coli JM109 on 2X YT agar
at a density of 20 to 30 plaques per 100mm plate. Individual phage particles
are obtained by selecting all the plaques on each plate for a total of 100 or
more, to minimize bias in the selection process. Each agar plug selected was
used to infect early log phase JM109 cells grown in 2X YT broth, and allowed to
amplify for 8 to 9 hours at 37ºC. Double-stranded replicative form DNA was
isolated from pelleted infected cells with Qiagen plasmid mini-spin columns
according to manufacturer's instructions. DNA sequences of the inserts were
obtained by direct PCR sequencing of the DNA using the recommended primer
sequence. The resulting data is analyzed using DNA2PRO. For more details, see
Rodi, D.J., Janes, R.W., Sanganee, H.J., Holton, R., Wallace, B.A. and
Makowski, L. (1999) Screening a library of phage-displayed peptides identifies
human Bcl-2 as a Taxol-binding protein, J. Mol. Biol. 285,197-204.
For a detailed protocol of these methods, please contact
Diane Rodi
.
To download our affinity selected peptide libraries, please click on the name
links!
ATP
|
name
|
marker
|
# peps
|
library
|
special conditions
|
|
atpa-12s
|
biotin
atp
|
446
|
NEB
12
|
standard conditions
|
|
atpc-12s
|
biotin
atp
|
89
|
NEB
12
|
prepanned phage on
SA plate
|
|
atpd-12s
|
biotin
atp
|
82
|
NEB
12
|
had SA in phage
solution during biopanning
|
|
atpeb-12s
|
biotin
atp
|
87
|
NEB
12
|
stayed on plate
after soluble ATP treatment
|
|
atpes-12s
|
biotin
atp
|
91
|
NEB
12
|
competed off plate
w/soluble ATP
|
|
gatp-12s
|
gamma
atp
|
360
|
NEB
12
|
standard conditions
|
|
satp-12s
|
Bodipy-fl
atp
|
86
|
NEB
12
|
standard conditions
|
|
gatp-c7s
|
gamma
atp
|
551
|
NEB
C7C
|
standard conditions
|
|
atpa-c7s
|
biotin
atp
|
556
|
NEB
C7C
|
standard conditions
|
GTP
|
name
|
marker
|
# peps
|
library
|
special
conditions
|
|
gtp-12s
|
bodipy-fl gtp
|
92
|
NEB
12
|
standard
conditions
|
GLUCOSE
|
name
|
marker
|
#
peps
|
library
|
special conditions
|
|
higlu-12s
|
biotin
GLUCOSE
|
91
|
NEB
12
|
re-do of gluc pan at
higher salt con.
|
|
loglu-12s
|
biotin
GLUCOSE
|
94
|
NEB
12
|
panned twice w/out
amp., at low then hi salt
|
GALACTOSE
|
name
|
marker
|
#
peps
|
library
|
special conditions
|
|
gal-12s
|
biotin
GALACTOSE
|
87
|
NEB
12
|
panned at high salt
|
TAXOL / TAXOTERE
|
name
|
marker
|
#
peps
|
library
|
|
TX-12s
|
biotin
taxol
|
69
|
NEB
12
|
|
TX-c7s
|
biotin
taxol
|
98
|
NEB
C7C
|
|
TXE-12s
|
biotin
taxotere
|
105
|
NEB
12
|
|
TXE-c7s
|
biotin
taxotere
|
100
|
NEB
C7C
|
STREPTAVIDIN
|
name
|
marker
|
#
peps
|
library
|
special
conditions
|
|
SA-12s
|
streptavidin
|
95
|
NEB
12
|
Pending |
|
SA-c7s
|
streptavidin
|
106
|
NEB
C7C
|
Pending |
VEGF
|
