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. Author manuscript; available in PMC: 2023 Jul 23.
Published in final edited form as: J Proteome Res. 2019 Apr 23;18(5):2270–2278. doi: 10.1021/acs.jproteome.9b00118

Figure 1.

Figure 1.

Flowchart outlining the workflow for the hyper-citrullinated library generation pipeline. A total of six tissues were isolated from three male wild-type mice. The tissues were snap-frozen, homogenized, and solubilized, and the total protein concentration was determined using a BCA protein assay kit. Step 1. Library building. In brief, each sample was split into two samples; one was treated with PAD cocktail (five isoforms, ratio 1:20) to cover all potential citrullinated targets. The corresponding second sample was treated with water under the same condition. Step 2. Digestion with LysC (ratio 1:20), followed by LC–MS/MS (TripleTOF 5600). Step 3. Postprocessing analysis with algorithm that examines the MS runs for the evidence of citrullinated site and Skyline validation.