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The application of westerns or immunoblotting techniques for assessing the composition, dynamics, and purity of protein extracts from plant material has become common practice. Open Preview See a Problem? The triple quadrupole mass spectrometers required are relatively affordable, further increasing the potential popularity of MRM for such approaches. This technique relies on the assumption that changes in abundance of an entire subcellular compartment can be represented by a small number of proteins. Detection is biased against very small, low abundance, or hydrophobic proteins using data-dependent acquisitions, particularly in complex samples, whilst heavily post-translationally modified proteins may never be detected. Estimation of relative abundance of cellular compartments from total protein extracts of 7-day old cell suspension cultures A,C or 4-week rosettes B,D. Given the history of using approaches like westerns and enzyme assays to assess organelle contributions in a sample, the MRM technique could be extended from the individual protein to the compartment level by designing suites of peptide transitions covering marker proteins for multiple subcellular compartments.
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