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

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 c...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2044; p. 81
Main Authors Rao, Aishwarya A, Mehta, Kanika, Gahoi, Nikita, Srivastava, Sanjeeva
Format Journal Article
LanguageEnglish
Published United States 2019
Subjects
Brain Neoplasms - cerebrospinal fluid
Brain Neoplasms - chemistry
Cerebrospinal Fluid Proteins - analysis
Cerebrospinal Fluid Proteins - chemistry
Cerebrospinal Fluid Proteins - isolation & purification
Chemical Fractionation - instrumentation
Chemical Fractionation - methods
Glioma - cerebrospinal fluid
Glioma - chemistry
Humans
Mass Spectrometry
Proteome - chemistry
Proteome - metabolism
Proteome - standards
Proteomics - methods
Proteomics - standards
Software
Staining and Labeling - methods
Tumor Microenvironment - genetics
Two-Dimensional Difference Gel Electrophoresis - methods
Workflow
Cyanine dyes
Sample fractionation
DIGE
Sample preparation
Mass spectrometric analysis
Internal standard
Reaction quenching
iTRAQ
Isobaric labeling
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Summary: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.
ISSN:1940-6029
DOI:10.1007/978-1-4939-9706-0_6