Biology Reference
In-Depth Information
Table 2
Summary of different proteomic methodologies used for soybean proteome analyses
Plant tissue
Protein extraction buffer
Protein solubilization/lysis buffer
Proteomic methodologies
Spot resolved
Reference
Roots
10 % TCA in acetone containing
0.07 % DTT, 1 % PVP
7 M urea, 2 M thiourea, 2 % CHAPS,
1 % DTT, 2 % ampholytes (pH 3-10)
IPG, 2-DE,
MALDI-TOF MS
>1,200
[
17
]
10 % TCA, 0.07 % 2-ME
8 M urea, 2 M thiourea, 5 % CHAPS,
2 mM tributylphosphine, ampholytes
(pH 3-10)
IPG, 2-DE,
nanoLC-MS/MS
439
[
16
]
0.5 M Tris-HCl (pH 8.3), 2 % NP-40,
20 mM MgCl
2
, 2 % 2-ME, 1 mM
PMSF, 1 % PVP/fractionation with
PEG 4000
8 M of urea, 1 % CHAPS, 0.5 % IPG
buffer pH 4-7, 20 mM DTT, BPB
IPG, 2-DE, MALDI-TOF
MS, ESI-MS/MS
~900
[
27
]
Root apex/
differentiated
root zone
Acetone containing 10 % TCA,
0.07 % DTT
9 M urea, 4 % CHAPS, 1 % DTT, 0.8 %
ampholytes (pH 3-10), 35 mM
Tris-HCl, 1 mM PMSF, 5 mM EDTA
IPG, 2-DE, DIGE
Labeling, CBB staining,
MALDI-TOF MS/MS
1630 Cy-dye-/
550 CBB
[
18
]
Root tips
10 % TCA, 0.07 % 2-ME in acetone
8 M urea, 2 M thiourea, 5 % CHAPS,
2 mM tributylphosphine
SDS-PAGE, ProQ
Diamond
phosphoprotein gel stain,
nanoLC-MS/MS
-
[
46
]
Root hairs
50 % phenol, 0.45 M sucrose, 5 mM
EDTA, 0.2 % 2-ME, 50 mM Tris-
HCl pH 8.8
8 M urea, 2 M thiourea, 2 % CHAPS,
2 % Triton X-100, 50 mM DTT,
2 mM TBP, 0.5 % ampholytes
IPG, 2-DE, MALDI-TOF
MS, Q-TOF-MS
-
[
37
]
Roots and
hypocotyls
Phosphate saline buffer (pH 7.6)
containing 65 mM
K
2
HPO
4
, 2.6 mM KH
2
PO
4
, 400 mM
NaCl and 3 mM NaN
3
followed by
10 % trichloro acetic acid
8.5 M urea, 2.5 M thiourea, 5 %
CHAPS, 100 mM dithiothreitol and
0.5 % ampholytes (pH
3.0-10.0/5.0-8.0)
IEF tube gel, 2-DE,
MALDI-TOF MS
235 (R)
330 (H)
340 (L)
[
33
]
Leaves
-
7 M urea, 0.2 M thiourea, 0.2 mM TBP,
0.4 % CHAPS, 0.2 % ampholytes (pH
3.0-10.0), 5 % PVP-40
