Abstract
The proteome data provided in this article were acquired from MCF7 breast cancer cells stimulated with insulin, and were generated by using a 2D-SCX (strong cation exchange)/RPLC (reversed phase liquid chromatography) separation protocol followed by tandem mass spectrometry (MS) detection. To facilitate data re-processing by more advanced search engines and the extraction of additional information from already existing files, both raw and processed data are provided. The sample preparation, data acquisition and processing protocols are described in detail. The raw data relate to work published in “Proteome profile of the MCF7 cancer cell line: a mass spectrometric evaluation” (Sarvaiya et al., 2006) [1] and are made available through the PRIDE (PRoteomics IDEntifications)/ProteomeXchange public repository with identifier PRIDE: PXD004051 (“2016 update of the PRIDE database and tools” (Vizcaino et al., 2016) [2]).
Highlights
The proteome data provided in this article were acquired from MCF7 breast cancer cells stimulated with insulin, and were generated by using a 2D-SCX/RPLC separation protocol followed by tandem mass spectrometry (MS) detection
Data are provided in this article and MS RAW and processed files have been deposited in the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PRIDE [2]: PXD004051 and http://dx.doi.org/DOI:10.6019/PXD004051
The data provided in this manuscript describe the proteome profile of insulin-stimulated MCF7 breast cancer cells
Summary
The cells were lysed by rocking with RIPA buffer supplemented with protease and phosphatase inhibitors for 2 h at 4 °C. The sample (16 mL injection containing 64 mg protein digest) was eluted from the SCX column in 16 fractions by a gradient consisting of: 100% A (0–5 min), 0–5% B (5–5.1 min), 5–20% B (5.1–35 min), 20–100% B (35–40 min), 100% B (40–50 min), 100 to 0% B (50–50.1 min) and 100% A (50.1–60 min). Reversed phase nano-LC separations were performed with home-built capillary columns (100 mm i.d. Â 12 cm fused silica capillaries) packed with Zorbax SB-C18 (5 mm) particles (Agilent). The Agilent 1100 micro-HPLC system was modified with a home-built split/splitless setup to allow for the generation of solvent gradients in the nanoliter/min flow regime. Each SCX fraction was loaded separately on the nano-LC column (40 mL), and eluted by an eluent gradient at $ 160–180 nL/min. The gradient consisted of: 0–10% B (0– 1 min), 10–45% B (1–95 min), 45–60% B (95–110 min), 60–100% B (110–115 min), 100%B (115–120 min), 100 to 0% B (120–121 min) and 100% A (121–150 min)
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