Assessment of ProteoExtract subcellular fractionation kit reveals limited and incomplete enrichment of nuclear subproteome from frozen liver and heart tissue |
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| Authors: | Christopher I. Murray Meagan Barrett Jennifer E. Van Eyk |
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| Affiliation: | 1. Department of Physiology, Queen's University, Kingston, Canada;2. Department of Biological Chemistry, Johns Hopkins University, Baltimore, MD, USA;3. Department of Medicine, Division of Cardiology, Johns Hopkins University, Baltimore, MD, USA |
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| Abstract: | The nuclear fraction of the ProteoExtract subcellular fractionation kit was assessed using frozen rat liver and heart tissue. Fractionation was evaluated by Western blot using protein markers for various subcellular compartments and followed up with LC/MS/MS analysis of the nuclear fractions. Of the proteins identified, nuclear proteins were in the minority (less than 15%) and there was poor representation of the various nuclear substructures when compared with liver nuclear isolations using a classical density‐based centrifugation protocol. The ProteoExtract kit demonstrated poor specificity for the nucleus and offers limited promise for proteomics investigations of the nuclear subproteome in frozen tissue samples. |
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| Keywords: | Cardiac muscle Fractionation Liver ProteoExtract |
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