After that, 200 μL NH4HCO3 solution (50 mM) was added to the filter and centrifuged at 14,000× g for 25 min. Finally, 1.25 μg of trypsin was added and the mixture incubated at 40 °C overnight [16 (link),17 (link)]. The digestion procedure was stopped by adding formic acid (10%), and digested peptides were collected by centrifuging at 14,000× g for 15 min. After desalting with the C18 tips, the collected peptides were vacuum-dried [17 (link),18 (link)]. After trypsin digestion, Ziptip C18 micropipette tips (Millipore) were used to purify the peptides prior to adding 0.1% formic acid for LC-MS/MS analysis.
C18 ziptip micropipette tips
The C18 ZipTip micropipette tips are a type of solid-phase extraction (SPE) tip used for sample preparation in analytical applications. The tips contain a sorbent material based on C18 reversed-phase chemistry, which allows for the selective extraction and concentration of analytes from complex samples prior to analysis.
5 protocols using c18 ziptip micropipette tips
Protein Reduction, Alkylation, and Trypsin Digestion
After that, 200 μL NH4HCO3 solution (50 mM) was added to the filter and centrifuged at 14,000× g for 25 min. Finally, 1.25 μg of trypsin was added and the mixture incubated at 40 °C overnight [16 (link),17 (link)]. The digestion procedure was stopped by adding formic acid (10%), and digested peptides were collected by centrifuging at 14,000× g for 15 min. After desalting with the C18 tips, the collected peptides were vacuum-dried [17 (link),18 (link)]. After trypsin digestion, Ziptip C18 micropipette tips (Millipore) were used to purify the peptides prior to adding 0.1% formic acid for LC-MS/MS analysis.
MALDI-TOF Mass Spectrometry Protocol
Quantitative Proteomics with TMT
Cell Lysis and Tryptic Digestion for Proteomics
MALDI-TOF-MS Peptide Detection Using SPE
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