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Application of 2D-DIGE and iTRAQ Workflows to Analyze CSF in Gliomas.

CEREBROSPINAL FLUID (CSF) PROTEOMICS: METHODS AND PROTOCOLS(2019)

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Abstract
Proteomics is an indispensable tool for disease biomarker discovery. It is widely used for the analysis of biological fluids such as cerebrospinal fluid (CSF), blood, and saliva, which further aids in our understanding of disease incidence and progression. CSF is often the biospecimen of choice in case of intracranial tumors, as rapid changes in the tumor microenvironment can be easily assessed due to its close proximity to the brain. On the contrary studies comprising of serum or plasma samples do not truly reflect the underlying molecular alterations due to the presence of protective blood-brain barrier. We have described in here the detailed workflows for two advanced proteomics techniques, namely, 2D-DIGE (two-dimensional difference in-gel electrophoresis) and iTRAQ (isobaric tag for relative and absolute quantitation), for CSF analysis. Both of these techniques are very sensitive and widely used for quantitative proteomics analysis.
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Key words
Cyanine dyes,DIGE,Internal standard,Isobaric labeling,Mass spectrometric analysis,Reaction quenching,Sample fractionation,Sample preparation,iTRAQ
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