Abstract
Due to post-sampling changes, caused by residual enzyme activity in the sample, levels of analytes can change from their in vivo levels so that they no longer accurately reflect conditions in the living system. The Stabilizor™ system accomplishes elimination of enzyme activity through heat-induced denaturation of enzymes by permanently altering the 3D protein structure of the enzymes. Heat stabilization can be introduced in the workflow either directly after sampling, with the instrument just next to where the sample is taken, or prior to sample homogenization and extraction, when samples are heat denatured directly from a frozen state. Initially, heat stabilization was developed to enable mass spectrometric analysis of neuropeptides. Heat stabilization has since been further developed and applied to a range of samples and downstream protein analysis techniques such as western blot, 2D gels and phosphorylation analysis with LC-MS.
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References
Sköld K, Alm H, Scholz B (2013) The impact of biosampling procedures on molecular data interpretation. Mol Cell Proteomics 12(6):1489–1501
Stingl C, Söderquist M, Karlsson O et al (2014) Uncovering effects of ex vivo protease activity during proteomics and peptidomics sample extraction in rat brain tissue by oxygen-18 labeling. J Proteome Res 13(6):2807–2817
Svensson M, Borén M, Sköld K et al (2009) Heat stabilization of the tissue proteome: a new technology for improved proteomics. J Proteome Res 8(2):974–981
Kultima K, Sköld K, Borén M (2011) Biomarkers of disease and post-mortem changes—heat stabilization, a necessary tool for measurement of protein regulation. J Proteomics 75(1):145–159
Ahmed MM, Gardiner KJ (2011) Preserving protein profiles in tissue samples: differing outcomes with and without heat stabilization. J Neurosci Methods 196(1):99–106
Robinson AA, Westbrook JA, English JA et al (2009) Assessing the use of thermal treatment to preserve the intact proteomes of post-mortem heart and brain tissue. Proteomics 9(19):4433–4444
Smejkal GB, Rivas-Morello C, Chang JH et al (2011) Thermal stabilization of tissues and the preservation of protein phosphorylation states for two-dimensional gel electrophoresis. Electrophoresis 32(16):2206–2215
Li X, Friedman BA, Roh MS, Jope RS (2005) Anesthesia and post-mortem interval profoundly influence the regulatory serine phosphorylation of glycogen synthase kinase-3 in mouse brain. J Neurochem 92:701–704
Borén M (2011) Methodology and technology for stabilization of specific states of signal transduction proteins. Methods Mol Biol 717:91–100
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Borén, M. (2015). Sample Preservation Through Heat Stabilization of Proteins: Principles and Examples. In: Posch, A. (eds) Proteomic Profiling. Methods in Molecular Biology, vol 1295. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2550-6_2
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DOI: https://doi.org/10.1007/978-1-4939-2550-6_2
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2549-0
Online ISBN: 978-1-4939-2550-6
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