Biology Reference
In-Depth Information
two ways to go about it: one is to remove the SDS via the classical
acetone/methanol precipitation, and the other is to dilute the SDS
from 3 to 0.1 % in the presence of another compatible surfactant,
like 0.5 % CHAPS, conditions that would be compatible with the
CPLL technology. There is a substantial amount of extra work
involved, yet the results have been outstanding: in the case of avo-
cado, the total number of unique gene products identifi ed amounted
to 1,012 proteins, of which 174 in common with the control,
untreated sample, 190 present only in the control, and 648 repre-
senting the new species detected via CPLLs of all combined (native
and denatured) eluates and likely representing low-abundance pro-
teins. In the case of banana, out of a total number of 1,131 identifi ed
proteins, the various captures (under native and denaturing condi-
tions) with the CPLL beads permitted the identifi cation of 849 pro-
teins, whereas the controls allowed identifying 452 proteins, 170
species being in common. Thus it is seen that with this double
extraction protocol, one can easily exceed the 1,000-species limit in
a single sweep, almost an anathema in plant proteomics. By compari-
son, in analysis of olive fruit pulp [
47
], where only native extraction
was applied, the booty was rather meagre: only 252 unique gene
products could be identifi ed. The readers are referred to these two
recent papers [
45
,
46
] for all practical details. For detailed basic
information and protocols sample treatment using various approaches
including recipes for plant extracts, the reader should refer to a
recently published topic as referenced [
48
].
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