Protein Biomarker Discovery Protocols
Depletion of high abundance proteins from plasma and iTRAQ labeling
In all 3 iTRAQ experiments, the number of plasma samples stated was pooled together according to their classification (i.e. normal, early gastric cancer, late gastric cancer or lung cancer) prior to depletion of high abundance proteins. For the first and second sets of iTRAQ experiment, a total of 35 mg of protein was pooled from each sample. The pooled samples were then delipidated by centrifugation at 130,000 x g at 4°C for 2 hr. The top transparent layer of plasma resulting from ultracentrifugation was collected and the total protein estimated using BCA assay (Pierce Biotechnology, IL, USA). Plasma samples were then subjected to depletion using lgY-12 spin column (Beckman Coulter, CA, USA) according to the manufacturer's protocol .
Disclaimer: Sciclips does not guarantee the authenticity or validity of the protocols listed in this section. These protocols are extracted "as is" from various sources, with minor modifications. Please refer to the respective citations, which are hyperlinked, for more details. The use of cited molecules, not limited to proteins, nucleic acids, compounds etc, in the protocols that are extracted from patents may be covered under patent rights. These molecules should be used for information purpose only.
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