Proteomics

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Evaluating cPILOT Performance on Orbitrap Instruments

ABSTRACT: A global cPILOT assay developed on an Orbitrap Velos instrument was transitioned to an Orbitrap Fusion Lumos instrument. Parameters such as the LC gradient, m/z isolation window, dynamic exclusion, targeted mass analyses, and SPS-N were optimized. The number of proteins identified on the Fusion Lumos (1848) without fractionation are close to those identified to the Orbitrap Velos (2199) with sample pre-fractionation. Additionally, including sample fractionation with the Fusion Lumos optimized parameters resulted in the identification of 4968 proteins, of which 1234 were quantified in each tissue type and genotype (N=5 or 6). Combined, the various experiments described herein informed of system-wide changes in brain, heart, and liver proteins from a late-stage amyloid precursor protein/presenilin-1 (APP/PS-1) human transgenic double knock – in mouse model of AD.

INSTRUMENT(S): Orbitrap Fusion Lumos, LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Heart, Brain, Liver

DISEASE(S): Alzheimer's Disease

SUBMITTER: Christina King  

LAB HEAD: Rena A.S. Robinson

PROVIDER: PXD012133 | Pride | 2021-09-08

REPOSITORIES: Pride

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Publications

Evaluating Combined Precursor Isotopic Labeling and Isobaric Tagging Performance on Orbitraps To Study the Peripheral Proteome of Alzheimer's Disease.

King Christina D CD   Robinson Renã A S RAS  

Analytical chemistry 20200127 4

Combined precursor isotopic labeling and isobaric tagging (cPILOT) is an enhanced multiplexing strategy currently capable of analyzing up to 24 samples simultaneously. This capability is especially helpful when studying multiple tissues and biological replicates in models of disease, such as Alzheimer's disease (AD). Here, cPILOT was used to study proteomes from heart, liver, and brain tissues in a late-stage amyloid precursor protein/presenilin-1 (APP/PS-1) human transgenic double-knock-in mous  ...[more]

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