TY  - BOOK
AU  - Goldman,A.R.
AU  - Beer,L.A.
AU  - Tang,H.Y.
AU  - Hembach,P.
AU  - Zayas-Bazan,D.
AU  - Speicher,D.W.
TI  - Proteome Analysis Using Gel-LC-MS/MS
N2  - This article describes processing of protein samples using 1D SDS gels prior to protease digestion for proteomics workflows that subsequently utilize reversed-phase nanocapillary ultra-high-pressure liquid chromatography (LC)coupled to tandem mass spectrometry (MS/MS). The resulting LC-MS/MS data are used to identify peptides and thereby infer proteins present in samples ranging from simple mixtures to very complex proteomes. Bottom-up proteome studies usually involve quantitative comparisons across several or many samples. For either situation, 1D SDS gels represent a simple, widely available technique that can be used to either fractionate complex proteomes or rapidly clean up low microgram samples with minimal losses. After gel separation and staining/destaining, appropriate gel slices are excised, and in-gel reduction, alkylation, and protease digestion are performed. Digests are then processed for LC-MS/MS analysis. Protocols are described for either sample fractionation with high-throughput processing of many samples or simple cleanup without fractionation. An optional strategy is to conduct in-solution reduction and alkylation prior to running gels, which is advantageous when a large number of samples will be separated into large numbers of fractions. Optimization of trypsin digestion parameters and comparison to in-solution protease digestion are also described
UR  - https://drive.google.com/file/d/1IqOjZ90eMUoQNK_q7FqiP1hTxwiLISXl/view?usp=drivesdk
ER  -