obtained within 15 min after collection. The collection tubes are
centrifuged for 15 min at 700 Â g at room temperature. The
plasma is separated from blood cells, aliquoted, and stored at
À80
C until analysis.
l
To obtain serum, blood samples are collected from the cubital
vein into blood collection tubes without any additives
[55]. Serum is obtained after the coagulation of blood for 1 h
at room temperature.
The collection tubes are centrifuged for 15 min at 700 Â g at
room temperature. Serum is separated from the clot, aliquoted, and
stored at À80
C until analysis.
Serum Sample Preparation
[57–59]
There are some authors that establish protocols to make a proteomics assay in serum samples without protein depletion or
pre-fractionation [59]. However, the major proteins can avoid
minor protein identification, which in most cases are the most
interesting proteins (Fig. 7).
1. Three aliquots of 30 μL of human serum are precipitated using
MeOH/CHCl 3 precipitation (see before for more details
about the precipitation protocol).
2. The pellet is resuspended in Milli-Q water and submitted to an
in-gel concentration and in-gel digestion (see next section).
Fig. 6 A representative workflow of the quantitative SWATH-MS proteomics of urine. Urine samples are
collected from human patients (control and disease groups) and centrifuged to remove particulates and cell
debris. After protein digestion and desalting, each sample is submitted to a LC-MS/MS analysis
SWATH-MS Protocols in Human Diseases
119
centrifuged for 15 min at 700 Â g at room temperature. The
plasma is separated from blood cells, aliquoted, and stored at
À80
C until analysis.
l
To obtain serum, blood samples are collected from the cubital
vein into blood collection tubes without any additives
[55]. Serum is obtained after the coagulation of blood for 1 h
at room temperature.
The collection tubes are centrifuged for 15 min at 700 Â g at
room temperature. Serum is separated from the clot, aliquoted, and
stored at À80
C until analysis.
Serum Sample Preparation
[57–59]
There are some authors that establish protocols to make a proteomics assay in serum samples without protein depletion or
pre-fractionation [59]. However, the major proteins can avoid
minor protein identification, which in most cases are the most
interesting proteins (Fig. 7).
1. Three aliquots of 30 μL of human serum are precipitated using
MeOH/CHCl 3 precipitation (see before for more details
about the precipitation protocol).
2. The pellet is resuspended in Milli-Q water and submitted to an
in-gel concentration and in-gel digestion (see next section).
Fig. 6 A representative workflow of the quantitative SWATH-MS proteomics of urine. Urine samples are
collected from human patients (control and disease groups) and centrifuged to remove particulates and cell
debris. After protein digestion and desalting, each sample is submitted to a LC-MS/MS analysis
SWATH-MS Protocols in Human Diseases
119