Label-free Protein Profiling
Regulated proteins can be Identified by relative quantitation of hundreds to thousands of proteins in a system of interest. The label-free approach is based on quantitation at MS1 levels and allows for a large number of biological and/or technical replicates to be included in the experimental design.
|Platform||Q-Exactive-Quadropole coupled to an Orbitrap analyser|
|Research Objectives||Relative quantitation of protein levels in perturbed systems|
This service provides a means to discover proteins, and consequently pathways, which may be of importance in biological response systems to elicitors such as disease, stress and drug treatment. Importantly, this discovery approach provides an unbiased means of finding novel leads for further downstream research.
Through client consultation, an applicable experimental approach will be designed. Once samples have been supplied, total protein concentration will be determined and the total protein amount equalised for each sample. Protein will thereafter be subject to tryptic digestion and fractionated by 1D or 2D gel electrophoresis. Once fractionated each sample will be subjected to MS analysis. The resulting data will be analysed and proteins identified using an appropriate database search tool. Relative protein levels will be quantified and a statistical analysis of identified and quantified proteins performed.
An analytical report will be provided that contains a comprehensive list of identified and regulated proteins as well as a statistical report assessing the quality of protein identification and quantification. Raw data files will also be made available.
- Protein Quantity: 5 µg – 2 mg (dependant on nature of sample prep)