Label-free Quantitative Proteomics
Introduction:
This service offers identification and quantification of the proteome in your complex samples without chemical modifications. The data generated by this service is a complete catalogue of the proteins present in your samples together with a statistical analysis result demonstrating the abundance of each identified protein in your samples. This service enables researchers to compare the proteome difference in their treated vs untreated, diseased vs normal, over-expressed vs control, gene silenced vs control, knockout vs wild type, mutant vs wild type groups, etc, with a relatively good accuracy. For better accuracy and reproducibility, please refer to our iTRAQ/TMT Labeling Quantification Service.
Your samples (several test groups and control group) will be digested with Trypsin and cleaned by C18 cartridges. To improve the sensitivity, we suggest the basic Reverse Phase LC (bRPLC) to further fractionate the cleaned sample into 96 fraction and then pooled to 24 fractions, which are subject to 24 Mass Spectrometry Analysis runs by the world's most advanced Orbitrap LC-MS/MS system.
There are 7 steps in a Label Free Omics Technologies quantitative proteomics project:
1) Sample preparation – Cell or tissue lysis, biofluid (plasma or serum) depletion; protein quantification on all samples.
2) Sample digestion – Reduction and alkylation followed by in-solution trypsin digestion and C18 cleaning before bRPLC.
3) 2D fractionation – bRPLC separation of the C18 cleaned samples into 96 fractions, and further pooled into 24 fractions based on the rule of avoiding co-elution.
4) Nano LC/MS/MS – 50 nL/min liquid chromatography to enable super high ionization efficiency for downstream Mass Spectrometry Analysis using the state-of-the-art Orbitrap Mass Spectrometer.
5) Protein profiling – Protein database searching with Mascot.
6) Data analysis – Data validation, visualization and quantification using commercial softwares and Omics Technologies's unique R packages and scripts.
7) Report generation – Report will be sent to you in Excel format as well as a summary in PDF format. We will also provide you any details you need for your papers' MATERIALS AND METHODS section. We will make sure you understand your result and help you with your paper writing with free follow-up services.
Sample types we accept:
1, 2D gel spots, SDS-PAGE bands
2, Cell Lysates and Tissue Lysates
3, Biofluids, such as plasma*, serum*, saliva, tear, etc.
*We provide High Abundance Protein Depletion Service to significantly (100-500 folds) increase the depth of your biofluid proteomics analysis by removing top abundant proteins from your samples. Read more for details.
4, FFPE slide/ FFPE extract
5, Customized sample types (please contact us to discuss)
Please refer to our Omics Technologies Proteomics Sample Preparation and Shipping Guidelines for instructions on how to prepare your samples with less contamination for Mass Spectrometry Analysis.
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Phone:
+1 888-206-0066 (TOLL FREE) -
Email:
info@omicstech.com