JRD235ITRAQ protein detection
ITRAQ protein detection ITRAQ protein detection brief introduction: Protein group Analysis (Proteomics) is a protein group (Proteome) analysis and detection, the protein group analysis first to separate mixed proteins, the main method of separation is protein bidirectional electrophoresis. The proteins that have been separated are identified by mass spectrometry.
The four isotopic labeling reagents were reported from 114, 115, 116, and 117, respectively, and relative molecular masss of 31, 30, 29, and 28, respectively. (Balance Group) and a peptide reactive group. Different reporting groups respectively match the corresponding balance groups, relative molecular masss are 145, namely the isobaric tag. Cracking the protein into a peptide, and then labeled with ITRAQ reagents. Since the ITRAQ reagent is equal, the different isotope is labeled with the same polypeptide, the molecular weight is the same, the molecular weight is the same, and the series mass spectrometry method is used to detect the precursor ion (Precursor ION) induction dissociation, product Ions are analyzed by second-level mass spectrometry. During the secondary mass spectrometry, the bond barrier between the report group, the mass balance group and the polypeptide reactive group, the mass balance group lost, producing a low quality (M / Z) report ion. Since the secondary mass spectrometry can analyze the reporting group of the relative molecular mass difference 1, the difference in ion intensity of different reporting groups represents the relative abundance of the polypeptide it labeled. At the same time, the amide bonds in the polypeptide are broken, forming a series of B ions and Y ions to obtain the mass number of the ionic fragment, and can identify the corresponding protein precursors through the database query and comparison.
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1. Genomics: Gene chip, SNP detection, DNA methylation detection, biological information analysis
[ 123] 2. Transcriptional group: microRNA chip, LNCRNA chip, expression spectrum chip, RNA-SEQ
3. Proteomics: 2D-DiGe, SiLAC , ITRAQ, TMT, Label-Free, MRM
5. Protein Detection: Western Blot, Co-IP EMSA, CHIP, Immunofluorescence, ELISA
6. Pathological testing: He dyeing, special dyeing, tissue sections, immunohistochemical, flow cytometry
7Metabolite: GC-MS, LC-MS, NMR 8. Cellular and animal model: primary cell, cell model, animal model construction
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