Protein mixtures were separated using (Two-dimensional gel electrophoresis (2D) and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE), the target protein were collected, and the peptide mixtures were extracted by in-gel digestion and later used for LC-MS/MS analysis to obtain protein information.
After the peptides are ionized in the mass spectrometer, the mass-to-charge ratio (m/z) of each peptide is analyzed by the mass spectrometer, and thus the relative molecular mass of each peptide is known. To obtain the sequence information of peptides, the mass spectrometer will select certain peptides for fragmentation and analysis to obtain secondary mass spectra. The search software compares the secondary mass spectrum information with the corresponding database to obtain the exact sequence of the peptide, and then splices the complete sequence of each protein, thus enabling the identification of the protein.
What We Provide
Creative Proteomics uses LC-MS/MS technology to be able to efficiently and accurately identify proteins in SDS-PAGE and 2-DE samples, and optimize the in-gel digestion method to significantly improve the success rate of protein mass spectrometry identification and peptide coverage.
Workflow
Application
soelectric Point Analy Identification of target proteins.
Protein mixture identification.
Technical Features
High sensitivity: identification of 2D sample with very low concentration.
High versatility: identification of SDS-PAGE sample prepared by IP, co-IP and pull-down method.
High throughput: tens to hundreds of proteins can be identified at a time.
Liquid phase coupled with mass spectrometry, automated operation, fast analysis and good separation.
Simple samples to detect easily.
Highly targeted to analyze easily.
Short project cycle time.
Platform
Q Exactive
LTQ Orbitrap Velos
6500 Q TRAP
Nano-LC
Reports
Experiment procedures
Parameters of liquid chromatography and mass spectrometer
MS raw data files
Peptide identifications and intensity
Protein identifications and intensity
Sample Type
2-D: clearly visible spots of coomassie bright blue staining or silver staining.
SDS-PAGE: clearly visible bands of coomassie bright blue staining or silver staining. single protein and purity >90%
Protein solution: single protein and purity >90%
Magnetic beads
IP product
Protein powder
Peptide powder
