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- Liquid phase method: Liquid phase system: M Class system (Waters), chromatographic column: EV-1106 chromatographic column, 15 cm x 150 μm, 1.
9 μm particle size, flow rate: 300 nL/min, gradient: 45 min
.
- Mass spectrometry method: Mass spectrometry: ZenoTOF™ 7600 system, acquisition mode: Zeno SWATH DIA, variable number of windows: 80 (m/z 400-903), primary accumulation time: 50 ms, secondary accumulation time: 18 ms, source Gas parameters: ion source voltage 3200 V, temperature 225°C, Gas1: 10 psi, CUR gas: 20 psi
.
- Data analysis: Zeno SWATH DIA data were processed by DIA-NN software, using self-built database and non-self-built library process, respectively.
The self-built database was the human cell lysate database previously established by OneOmics
.
Main results:
1.
Quantitative protein identification and quantification at low injection volumes
Figure 1.
Proteins in different low-injection samples (left).
Proteins in different low-injection samples (left).
Identification and quantification of peptides (right)
2.Comparison of the number of protein identifications when using the self-built database and the non-self-built database
Figure 2.
Using a database versus a non-database analysis process
Comparison of protein identification numbers
3.Zeno SWATH IDA Process Stability Test
Figure 3.
Comparison of different times with independently diluted K562 samples
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