AI Article Synopsis
- Researchers conducted an assessment of peptides from the mouse hippocampus by enzymatically digesting a protein extract and fractionating it via isoelectric focusing (IEF).
- The analysis revealed that samples with multiple individual fractions led to higher protein identification rates and improved quantification compared to unfractionated samples.
- The study concluded that while prefractionation enhances proteome resolution, the benefits plateau after a certain point, suggesting a strategic approach to pooling fractions can optimize costs and data effectiveness.
Article Abstract
An assessment of fractionated mouse hippocampal peptides was conducted. Protein extract from a single mouse hippocampus was enzymatically digested and fractionated by IEF. Aliquots of fractions were pooled into fewer, more complex samples. The unfractionated lysate, fractions, and pooled fractions were subjected to LC-MS/MS analysis. Samples consisting of many individual fractions had more protein identifications, greater protein sequence coverage, and quantified proteins with more spectral counts than protein extract that was unfractionated or pooled into fewer LC-MS/MS samples. Additionally, prefractionation reduced the median CV for spectral counts as much as 33%. However, the relative gain in proteome resolution was found to saturate with increasing fractionation extent. This study demonstrates how prefractionation by offline IEF can improve the resolution of proteomic investigations of the mouse hippocampus, and that a data-driven pooling methodology can reduce excessive and cost-ineffective fractionation.
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| http://dx.doi.org/10.1002/elps.201600076 | DOI Listing |
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