kD

1
A
pI
7.0
5.0
3.0
10.0
4.0
6.0
8.0
9.0
kD
5 DAF
9 DAF
7 DAF
116
97
84
66
55
45
36
24
29
20
7.0
5.0
3.0
4.0
6.0
8.0
9.0
10.0
pI
kD
11 DAF
13 DAF
Reference
116
97
84
66
55
45
36
29
24
20
B
pI
kD
6.0
5.0
4.0
7.0
5 DAF
7 DAF
9 DAF
116
97
84
66
55
45
36
29
24
20
pI
6.0
5.0
4.0
7.0
kD
11 DAF
13 DAF
Reference
116
97
84
66
55
45
36
29
24
20
Supplemental Figure 1. Two-dimensional
electrophoretic (2-DE) analysis of proteins (50
?g) isolated from immature seeds of 5, 7, 9, 11,
or 13 days after flowering (DAF) and labeled with
n-hydroxysuccinimide-activated Cy5.
Approximately 200 seeds from each of the five
individual stages were pooled in biological
triplicate and proteins isolated for two
analytical and two reference gels (pH 3-10 and
4-7) per biological replicate. A, 2-DE analysis
of developing seed proteins using pH 3 to 10
wide-range IPG strips. B, 2-DE analysis of
developing seed proteins using pH 4 to 7
medium-range IPG strips. Gels are representative
of the three biological replicates that were
analyzed. The reference gel is a composite of
all five stages obtained by pooling of 10 mg of
protein from all five seed stages prior to
labeling.