In this approach, a pair of protein samples are differentially
labeled. And the combined sample pair is first separated by SDS-PAGE
followed by coomassie staining. The entire gel lane containing the
labeled samples are sliced in the way that all abundant protein gel
bands are separately excised from the rest of gel slices. Typically
20-30 gel bands/slices will be cut out to cover an entire gel lane. Each
gel band is digested with a proteolytic enzyme and resulted peptides
from each gel band is analyzed by LC-MS to identify differential
peptides and then by LC-MS/MS to identify differential proteins. The
data from 20-30 gel bands is then integrated and validated.
The final report will includes all protein differentials identified
and ratio of concentration of differential proteins in the sample pair.
Deep
Analysis
