Nanoelectrospray nESI-IMS-MS experiments were carried out on a Synapt HDMS instrument (Waters Corp., Wilmslow, Manchester, UK). Samples of MEDI1912 and MEDI1912_STT were dialysed at 1 mg/mL into 150 mM ammonium acetate, pH 6 using dialysis buttons (Hampton Research Corp., Aliso Viejo, CA, USA). nESI-MS experiments were conducted in positive ion mode with samples being introduced using capillaries made in-house. The following instrument settings: capillary voltage 1.5 kV, sample cone 30 V, extraction cone 4 V, source temperature 80 °C, backing pressure 5.0 mBar, trap voltage 40 V, trap/transfer gas flow 1.5 mL/min, IMS gas flow 20 mL/min, IMS ramped wave height 5–30 V, travelling wave speed 300 ms. The m/z scale was calibrated using aqueous caesium iodide (CsI) cluster ions.
Synapt hdms instrument
Manufactured by Waters Corporation
Sourced in United Kingdom
The Synapt HDMS instrument is a high-performance mass spectrometry system designed for advanced analytical applications. It utilizes traveling wave ion mobility spectrometry (TWIMS) technology to separate and analyze complex samples. The Synapt HDMS provides high-resolution mass analysis and separation of ions based on their size, shape, and charge-to-mass ratio.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using synapt hdms instrument
1
MEDI1912 and MEDI1912_STT nESI-IMS-MS analysis
2
Optimizing Peptide Fragmentation and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peptide mixtures (both derivatized and underivatized) were diluted to a final concentration of 1 pmol/μL using 50% (v/v) acetonitrile in water containing 0.1% (v/v) formic acid, and infused into a Synapt HDMS instrument (Waters, Manchester, UK) using gold-coated nanospray emitter tips (Proxeon, Odense, Denmark). The capillary voltage, cone voltage, and source temperature were set at 2.3 kV, 40 V, and 80°C, respectively. The m/z of interest was mass selected by the quadrupole mass analyzer prior to being subjected to CID in the Trap collision cell. The collision energy applied in this region was manually tuned to give optimal peptide fragmentation, such that the precursor ion abundance was ~10% that of the base peak.
Further analysis of the peptide FGERALK (1 pmol/μL in 50% (v/v) acetonitrile in water containing 0.1% (v/v) formic acid was conducted by direct infusion nano-electrospray ionization (nESI) (5 μL/min) using an amaZon Ion Trap (Bruker, Bremen, Germany). Mass selection at m/z 410.7 corresponding to that of the peptide of interest was performed, followed by CID with the fragmentation amplitude set to 0.35 eV, with data collected between m/z 150 and 1800.
Further analysis of the peptide FGERALK (1 pmol/μL in 50% (v/v) acetonitrile in water containing 0.1% (v/v) formic acid was conducted by direct infusion nano-electrospray ionization (nESI) (5 μL/min) using an amaZon Ion Trap (Bruker, Bremen, Germany). Mass selection at m/z 410.7 corresponding to that of the peptide of interest was performed, followed by CID with the fragmentation amplitude set to 0.35 eV, with data collected between m/z 150 and 1800.
3
FOXM1 Mass Spectrometry Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mass spectra were recorded on a Synapt HDMS instrument (Waters UK Ltd., Manchester, UK). The concentration of FOXM1 was 4 μmol dm−3 while the final concentration of compounds after adding to the protein was 50 μmol dm−3. All measurements were carried out in a positive ion mode. Spraying conditions were: capillary voltage 1.7–1.8 kV, a cone voltage of 40–80 V, the source temperature 20°C, trap collision energy 12.0 V and transfer collision energy 12.0 V with trap and transfer pressure 5.31 × 10−2 mbar, IMS pressure 5.02 × 10−1 mbar, TOF analyser pressure 1.2 × 10−6 mbar. External calibration of the spectra was achieved using caesium iodide at 100 mg ml−1 in water. Data acquisition and processing were performed using Micromass MassLynx 4.1.
4
FOXM1 Mass Spectrometry Characterization
5
Native MS of Ctf4 with Dna2 and Tof2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In preparation for non-denaturing nano-electrospray ionization mass spectrometry (native mass spectrometry), Ctf4 471–927 was subjected to two successive rounds of buffer exchange into 500 mM ammonium acetate using illustra NAP-5 columns (GE Healthcare). Following buffer exchange, a 5-fold or 10-fold molar excess of Dna2 peptide 207- SLRNIDDILDDIEGDLT -223 or Tof2 peptide 497- SHAKDVKIQETIRKLNRFKPT -517 solubilized in 500 mM ammonium acetate was mixed with Ctf4CTD at a final protein concentration of 100 μM and incubated for a minimum of 30 min. Native mass spectra were recorded on a Synapt HDMS instrument (Waters) and calibrated using caesium iodide (100 mg ml−1) as described previously (Hernández and Robinson, 2007 (link), Simon et al., 2014 (link)).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!