Experimental Details - HuPA_00707
Experiment type Mass spectrometry
Short description Quantitative proteomic analysis of brain tissue proteome in toxoplasma encephalitis
Experimental description Frontal cortex brain tissues obtained from 5 toxoplasma encephalitis patients and 5 controls were used for protein extraction. Protein amount was normalized on 10% SDS- PAGE, equal amount of protein from 5 patients and 5 controls were pooled separately and subjected to reduction, alkylation and trypsin digestion. iTRAQ labelling of samples were carried out in replicates. Peptides from control samples were labelled with iTRAQ labels leading to the reporter ions of 114 and 115 whereas peptides from patient samples were labelled with iTRAQ labels leading to the reporter ions of 116 and 117. Labelled peptides were then pooled and subjected to SCX fractionation and obtained 21 fractions, which were subsequently subjected to high-resolution Fourier transform mass spectrometry (LTQ-Orbitrap Velos).
Principal Investigator's NameKeshava Prasad
TitleFaculty Scientist
AddressInstitute of Bioinformatics
Discoverer 7th Floor, ITPB
Bangalore, Karnataka 560066
Published/Unpublished Unpublished
Journal name Not applicable
PubMed ID Not applicable
Sample source
Tissue: Frontal cortex [eVOC:EV:0100167]
Cell line:
Source organism Homo sapiens [Taxonomy:9606]
Labeling technique iTRAQ
Protease used Trypsin
Is the sample from in gel No
Reduction and Alkylation Reduction of samples was carried out using Dithiothreitol followed by alkylation using iodoacetamide
Mass spectrometer
Instrument: LTQ Orbitrap Velos
Vendor: Thermo Scientific
Ionization method Nano ESI
Fragmentation method HCD
Mass tolerance used for database searching (MS) 20ppm
Mass tolerance used for database searching (MS/MS) 0.1 Da
Database used for searching Refseq 52
Search engine used Sequest
Download MS raw dataset  
Download processed files  

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