MS was performed on a Q Ex active Ultra-High Mass Range (UHMR) from ThermoFisher Scientific, modified with a custom reverse entry ion source (REIS) and 1.5 m drift tube. A Synapt G1 HDMS instrument with a 32-k RF generator from Waters Corporation was also used for membrane and soluble protein measurements. Nanoelectrospray ionization was performed as previously described.19 (link) UHMR parameters applied for AmtB included 250 °C capillary temperature, 1.40 kV spray voltage, 200 maximum inject time, resolution set to 6250 without averaging, 50 eV in-source CID, 40 CE, trapping gas pressure of 5. Ion transfer was set to high m/z mode, with in-source desolvation voltage set to −180 V.
Protocol detail
High-Resolution Mass Spectrometry for Membrane Proteins
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Corresponding Organization : Texas A&M University
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Variable analysis
independent variables
- UHMR parameters applied for AmtB
- Membrane and soluble protein measurements
- Capillary temperature: 250 °C
- Spray voltage: 1.40 kV
- Maximum inject time: 200
- Resolution: 6250 without averaging
- In-source CID: 50 eV
- Collision energy: 40 CE
- Trapping gas pressure: 5
- Ion transfer mode: high m/z
- In-source desolvation voltage: -180 V
- Positive control: Not mentioned
- Negative control: Not mentioned
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