Combination of 2DE and LC for Plant Proteomics Analysis - Plant Proteomics: Methods and Protocols - page 129

Sample
wheat embryos
(liquid chromatography)

First dimenssion
(Chromatophocusing
(HPCF))

44 % Area 25 % Area 31 % Area

9

0.12

0.10 8

0.08

7

0.06

6

0.04

5

0.02

4

0.00

3

0 25 50 75 100 125 150 175 200

Time (min)

Secon dimenssion
(High Performance Reversed Phase (RPRP))
p I

7.9 7.6 7.3 7.0 6.7 6.4 6.1 5.8 5.5 5.2 4.9 4.6 4.3 3.8

25

20

15

10

5

F10 F11 F12 F13 F14 F15 F16 F17 F18 F19 F20 F21 F22 F23 F24

Fig. 2 Liquid chromatography Proteomelab PF 2D. The fi rst step, chromatofocusing of total wheat embryo
proteins in a pH gradient. X -axis is retention time in the column, left y -axis pH, and right y -axis absorbance and
the second step, high-performance reversed phase. The fractions are separated in two dimensions: pH
gradient 8.0-4.0 and hydrophobicity. Finally were represented ProteomeLab PF 2D virtual gel. There are a total
of 15 fractions in the gradient and were detected 855 bands with a DeltaVue software. In the y -axis the reten-
tion time in minutes in the retention column is represented, in the upper x -axis the pH gradient, and in the
lower x -axis the fraction number