Our results highlighted proteins that were more prominent in one breed compared to the other (Table 1). For instance, a fatty acid-binding protein was 30% more abundant in Holstein-Friesian milk than Jersey milk. Samples were diluted with the loading buffer (0.5 M DTT added to 4X NuPAGE LDS Sample Buffer, Life Technologies) to reach a final 20 μL volume and heated at 70°C for 10 min. Placed into a systems biology context, method C (methanol/chloroform) is highly advantageous as it allows the recovery of polar, non-polar metabolites and proteins in one step. Для установки, подключения и обслуживания электрических приборов и/или цепей обращайтесь только к профессионалам, прошедшим cоответствующее обучение и имеющим допуск к работам. The recent sequencing of Bos taurus genome (Bovine Genome Sequencing and Analysis Consortium, 2009) paved the way for omics studies, particularly proteomics which heavily relies on gene model annotations for accurate protein identification. Within methods, cow breeds did not cluster together; it was evident within method A (urea) where Holstein-Friesian and Jersey breeds bear two different shades of colors that seldom mix.
Anal. Biochem. 150, 76–85. doi: 10.1016/0003-2697(85)90442-7 PubMed Abstract | CrossRef Full Text | Google Scholar Taylor, A. G., and Savage, C. (2006). Fatty acid composition of New Zealand green-lipped mussels, Perna canaliculus: Implications for harvesting for n-3 extracts. Chromatographic peaks were detected using a 1 min summation window and a curvature-based peak detection method. Method A (urea), a simple dilution of milk into an urea-based buffer, yielded the greatest number of unique protein accessions. The electrospray voltage was set at 2.2 kV, and the heated capillary was set at 280°C. Full MS scans were acquired in the Orbitrap Fourier Transform (FT) mass analyser over a normal range of 300–2000 m/z with 60,000 resolution in profile mode. MS/MS spectra were acquired in data-dependent mode. Funding This work was funded by Department of Economic Development, Jobs, Transport, and Resources. Two full cream milk samples were collected from bulk tanks containing the milk of the whole herd of Holstein-Friesian or Jersey cows milked on that particular day.
The subsequent clean-up, nLC-MS/MS, database search steps were rigorously performed as described above in Sections Tryptic Digest Cleaning, Assay and Dilution, Nano-Liquid Chromatography (nLC)-electrospray ionization (ESI) tandem MS (MS/MS) analyses, and Database Search for Protein Identification. Proteomics 4, 2094–2100. doi: 10.1002/pmic.200300723 PubMed Abstract | CrossRef Full Text | Google Scholar Hunt, J. M., and Tuder, R. (2012). Alpha 1 anti-trypsin: one protein, many functions. Данная таблица приведена для общей информации. Не забудьте ознакомиться с приложенной к устройству схемой. The original Bligh-Dyer procedure was subsequently modified by substituting water with a 8% NaCl solution (Taylor and Savage, 2006), thus blocking the binding of some acidic lipids to denatured lipids. The tubes were centrifuged at 13,000 rpm for 5 min at room temperature. Chromatograms were then aligned using a Pairwise Alignment Based Tree scheme with a 50 scan interval.