Advances in mass spectrometry and data analysis have provided a new paradigm to comprehensively investigate protein expression in complex biological systems. Cytomics have been providing mass spectrometry services for the analysis of proteins and proteomes since 2015 and today we make use of highly sensitive and accurate quantitative MS instrumentation and platforms to comprehensive survey even the most complex of biological systems. We can study alterations to key biological pathways which might be brought about by changes in disease state, condition, treatment or a stimulus. We access to this mass spectrometry platform and work closely with researchers in industry, academia and government organizations to support multidisciplinary projects which span the chemical and biological sciences, biomedicine and health, food and agriculture, and microbiology.
ThermoFisher Orbitrap platforms with nanoscale liquid chromatography systems for all projects. This is industry leading instrumentation employed by many academic thought-leaders.
Q Exactive HF-X hybrid quadrupole-Orbitrap mass spectrometers couples a high field Thermo Scientific™ Orbitrap™ mass analyzer with a high capacity transfer tube and electrodynamic ion funnel that maximize ion loading and transmit ions over a broad mass range. As a result, the instrument delivers a bright, focused and ion-rich beam, which translates to high sensitivity and excellent stability in analysis quality from run to run. Smart instrument control features match the superior ion transmission with increased acquisition speed and even more sophisticated data-dependent acquisition and analysis strategies to meet data quality needs at every stage of your work, from discovery to verification and quantitation
Advanced Quadrupole technology featuring Thermo Scientific segmented HyperQuad
Ultra-high-field Orbitrap analyzer, which doubles its speed and resolution resolving power up to 240,000
Maximum scan speed 40 Hz
Spectral multiplexing for enhanced duty cycle
High-capacity transfer tube and electrodynamic ion funnel source for increased sensitivity
Advanced Active Beam Guide intelligent ion beam management for high flux ion sources
The raw data files were processed using Proteome Discoverer (Version 220.127.116.11, Thermo Scientific). The data was searched using search engines SequestHT. We also use MaxQuant, Perseus and Skyline packages for data processing and review.
Type of services offered
A peptide centric (bottom up) approach using nanoLC-MS/MS and multiple modes of fragmentation (CID, HCD, ECD) to identify the amino acid sequences of peptides. Samples from solution or polyacrylamide gels are digested using trypsin (alternative enzymes available); peptides are purified, concentrated, and loaded onto a nanoLC C18 analytical column.
Intact Mass Measurement
An intact mass measurement can be used to provide confirm the sequence of a purified protein or to evaluate the binding of a ligand to a purified protein. In our experience, intact mass measurements are a invaluable in the determination of an unknown antibody sequence.
Protein Mapping - Protein Sequence Confirmation
Protein Mapping enables the identification of the entire sequence of a protein by using three enzymes for digestion. This service is ideal for identifying regions of interest inaccessible to trypsin alone, confirming a primary sequence or finding truncations. The basic experimental workflow is presented here
Relative Qualitative and Quantitative approaches indicating protein levels within the same sample as well as differential levels among samples. A variety of approaches exist within this category. Modern proteomics dictates that protein identification is only part of the puzzle, and increasingly it is necessary to understand the expression profiles of protein(s) within the overall context of the cell. Multiple solutions exist, some more accurate, robust and applicable than others. Our lab has experience with several methodologies including.
Tandem Mass Tags (TMT)
Parallel reaction monitoring (PRM)
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