Proteome Profiling
Introduction:
This is one of the most popular services in Omics Technologies Proteomics Services, and our service allows you to take full advantage of our most advanced Orbitrap Mass Spectrometers to identify your complete catalogue of the proteins present in your complex biological samples, such as cell lysates, tissue lysates, serum, plasma and other complicated samples.
Proteins in your samples are digested with a Omics Technologies unique combination of proteases, Trypsin and LysC. 2D fractionation is performed to further fractionate your samples into appropriate number of fractiontions according to your samples' complexicity. Peptides in total sample or each fraction are analyzed by world's most advanced Orbitrap LC-MS/MS. Identified peptides from your samples/fractions are matched to protein sequence database. 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.
There are typically 9 steps in a Omics Technologies Proteome Profiling 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.
3) Sample pooling – Equal amounts (according to total protein quantification) of labeled samples are pooled together to allow unbiased downstream purification and Mass Spectrometry Analysis.
4) SCX cleaning – Ion-exchange to remove excessive chemicals and fractionation of pooled sample through off-line SCX HPLC.
5) 2D fractionation – bRPLC separation of the SCX cleaned samples into 96 fractions, and further pooled into 24 fractions according to MyProt™-iPooling algorithm (based on peptide hydrophobicity, IEP, etc.)
6) 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. 24 fractions will be analyzed in 24 Mass Spectrometry runs.
7) Protein profiling – Protein database searching with Mascot, Sequest HT, PEAKS de novo approaches, etc.
8) Data analysis – Data validation, visualization and quantification using commercial softwares and Omics Technologies's unique R packages and scripts.
9) 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