Approximately 0.5 μg PR3 of each variant was purified using a Poros R1, 50 μm (Applied Biosystems, Carlsbad, CA, USA) microcolumn custom-made in a constricted GELoader tip (Eppendorff, Hamburg, Germany). The column was washed in 100% MeCN and equilibrated with 0.1% TFA prior to sample loading in 20 μL 0.1% TFA. After washing twice with 20 μL 0.1% TFA, the sample was eluted with 0.5 μL 20 mg/mL sinapinic acid in 0.1% TFA, 70% MeCN directly onto a dried droplet of 20 mg/mL sinapinic acid in 100% acetone placed on a metal target. MS was performed in a linear mode on a Bruker ultrafleXtreme MALDI instrument (Bruker Corporation, Billerica, MA, USA).
Ultraflextreme maldi instrument
Manufactured by Bruker
Sourced in United States, Germany
The UltrafleXtreme MALDI instrument is a high-performance mass spectrometry system designed for advanced analytical applications. It features a matrix-assisted laser desorption/ionization (MALDI) ion source and provides accurate mass measurements and high-resolution data acquisition.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ultraflextreme maldi instrument
1
PR3 Variant Purification and MALDI-TOF Analysis
2
Time-of-Flight Secondary Ion Mass Spectrometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
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A TOF.SIMS 5 mass spectrometer (ION-TOF,
Münster, Germany) was used for the time-of-flight secondary
ion mass spectrometry (TOF-SIMS) analyses. A pulsed 25 keV Bi3+ primary ion source was used as the analysis beam
(pulse width = 23 ns, mass resolution m/Δm = 5000). For the high lateral resolution imaging, pulse
width = 100 ns and mass resolution m/Δm = 500 were used. The primary ion dose was kept below the
static limit (1013 primary ions/cm2). Data was
collected in the positive ion mode.
MALDI data was acquired
on an UltrafleXtreme MALDI instrument (Bruker Daltonik GmbH, Germany)
equipped with a Nd:YAG smartbeam II laser (355 nm). The instrument
was operated in the reflectron mode. Positive ions were detected from
300 Da to 3 kDa. The laser power was optimized on the sample before
the data acquisition. For the MTF analysis, the “small”
laser focus setting (20 μm spot size) was used at a 50 μm
step size. 1000 laser shots were acquired at each shot at a 1 kHz
repetition rate.
Münster, Germany) was used for the time-of-flight secondary
ion mass spectrometry (TOF-SIMS) analyses. A pulsed 25 keV Bi3+ primary ion source was used as the analysis beam
(pulse width = 23 ns, mass resolution m/Δm = 5000). For the high lateral resolution imaging, pulse
width = 100 ns and mass resolution m/Δm = 500 were used. The primary ion dose was kept below the
static limit (1013 primary ions/cm2). Data was
collected in the positive ion mode.
MALDI data was acquired
on an UltrafleXtreme MALDI instrument (Bruker Daltonik GmbH, Germany)
equipped with a Nd:YAG smartbeam II laser (355 nm). The instrument
was operated in the reflectron mode. Positive ions were detected from
300 Da to 3 kDa. The laser power was optimized on the sample before
the data acquisition. For the MTF analysis, the “small”
laser focus setting (20 μm spot size) was used at a 50 μm
step size. 1000 laser shots were acquired at each shot at a 1 kHz
repetition rate.
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