Enhanced Sample Multiplexing for Nitrotyrosine-Modified Proteins Using Combined Precursor Isotopic Labeling and Isobaric Tagging

2012 ◽  
Vol 84 (11) ◽  
pp. 4677-4686 ◽  
Author(s):  
Renã A. S. Robinson ◽  
Adam R. Evans
Keyword(s):  
Isotopic Labeling ◽  
Modified Proteins ◽  
Isobaric Tagging

scholarly journals Automation of On-Resin Enrichment of S-Nitrosylated Proteins for Oxidized Cysteine-Selective cPILOT

2021 ◽  
Vol 11 ◽  
Author(s):  
Andrew Pumford ◽  
Albert Arul ◽  
Katarena Ford ◽  
Renã Robinson
Keyword(s):  
Quantitative Proteomics ◽  
Experimental Error ◽  
Normal Aging ◽  
Positive Pressure ◽  
Post Translational Modification ◽  
Liquid Handling ◽  
Sample Throughput ◽  
Proteomics Approach ◽  
Modified Proteins ◽  
Isobaric Tagging

S-Nitrosylation (SNO) is a cysteine post-translational modification that increases with normal aging and is present in Alzheimer’s disease and other aging-related illnesses. Detection of SNO-modified proteins can be challenging; however, we previously developed a robust quantitative proteomics approach termed “Oxidized Cysteine-Selective combined precursor isobaric labeling and isobaric tagging (OxcyscPILOT)” that allows for detection of endogenous SNO-modified proteins. OxcyscPILOT involves enrichment of SNO-modified proteins using a thiol-based resin. This enrichment is performed manually, and wash stages with the resin require numerous steps and buffer reagents. The goal of this study is to transfer the manual protocol to an automated liquid handler system in order to reduce wash steps, increase sample throughput, and minimize experimental error. In order to accomplish this, we evaluated the Biomek i7 liquid handler automated workstation and a Positive Pressure ALP (PPA) apparatus as a means to conduct automated on resin enrichment. Our findings provide starting pressure conditions for the use of PPA in an automated OxcyscPILOT proteomics workflow that could be transferred to other robotic liquid handling systems.

scholarly journals Increased N,N-Dimethyl Leucine Isobaric Tag Multiplexing by a Combined Precursor Isotopic Labeling and Isobaric Tagging Approach

2018 ◽  
Vol 90 (18) ◽  
pp. 10664-10669 ◽  
Author(s):  
Dustin C. Frost ◽  
Clayton J. Rust ◽  
Renã A. S. Robinson ◽  
Lingjun Li
Keyword(s):  
Isotopic Labeling ◽  
Isobaric Tag ◽  
Isobaric Tagging

scholarly journals Enhanced Sample Multiplexing of Tissues Using Combined Precursor Isotopic Labeling and Isobaric Tagging (cPILOT)

10.3791/55406 ◽  
2017 ◽  
Author(s):  
Christina D. King ◽  
Joseph D. Dudenhoeffer ◽  
Liqing Gu ◽  
Adam R. Evans ◽  
Renã A. S. Robinson
Keyword(s):  
Isotopic Labeling ◽  
Isobaric Tagging

scholarly journals High-throughput endogenous measurement of S-nitrosylation in Alzheimer's disease using oxidized cysteine-selective cPILOT

The Analyst ◽  
2016 ◽  
Vol 141 (12) ◽  
pp. 3904-3915 ◽  
Author(s):  
Liqing Gu ◽  
Renã A. S. Robinson
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Animal Models ◽  
High Throughput ◽  
Isotopic Labeling ◽  
Biological Tissues ◽  
Animal Models Of Disease ◽  
Isobaric Tagging ◽  
Models Of Disease

Combined precursor isotopic labeling and isobaric tagging (cPILOT) technique enables enhanced multiplexing quantification of S-nitrosylation in biological tissues from animal models of disease.

Accumulation of 4-hydroxy-2-nonenal-modified proteins in the white matter damage in chronic cerebral hypo-perfusion rat mode

2005 ◽  
Vol 25 (1_suppl) ◽  
pp. S262-S262
Author(s):  
Terubumi Watanabe ◽  
Yoshiko Yanagi ◽  
Takao Urabe ◽  
Yoshikuni Mizuno
Keyword(s):  
White Matter ◽  
White Matter Damage ◽  
Modified Proteins
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