Samples from immunoprecipitates were reduced, alkylated, and loaded onto SDS-polyacrylamide gel electrophoresis gels to remove LC-MS incompatible reagents. Gel plugs were excised, destained, and subjected to proteolytic digestion with trypsin and resulting peptides extracted and desalted, as previously described26 (link). Aliquots of the peptides were analyzed with LC-MS using a 60 min gradient on an EASY nLC 1000 coupled to a ThermoFisher Scientific Orbitrap Elite Hybrid Ion Trap Mass Spectrometer. The data were searched against a UniProt human database using Sequest within Proteome Discoverer. The results were filtered to only include proteins identified by at least two peptides. Protein quantitation was preformed using the ratios of PSMs in DAP5, eIF4GI, and eIF4GII AP over the PSMs in the control AP. In addition, the data were analyzed using the SAINT algorithm27 (link), including experiments 52–54 from the crapome.org database28 (link) as additional controls.
Orbitrap elite hybrid ion trap mass spectrometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Orbitrap Elite Hybrid Ion Trap Mass Spectrometer is a high-resolution, high-accuracy mass spectrometry platform designed for advanced analytical applications. It combines the Orbitrap mass analyzer with a linear ion trap, providing exceptional mass resolution, mass accuracy, and sensitivity.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using orbitrap elite hybrid ion trap mass spectrometer
1
Mass Spectrometry-Based Protein Identification
2
Nanoflow LC-MS/MS Analysis of Microbiome
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All samples were run on Eksigent 425 nanoHPLC connected to Orbitrap Elite™ Hybrid Ion Trap Mass Spectrometer [Thermo-Fisher Scientific, Waltham, MA, USA] with a 240 min gradient. Peptides were separated with an in-house made column (75 μm i.d. × 15 cm) packed with reverse phase beads [1.9 μm/120 Å ReproSil-Pur C18 resin, Dr. Maisch GmbH]. The samples were loaded at 5% buffer A (0.1% formic acid in H2O) and analyzed by a gradient from 5 to 30% (v/v) buffer B (0.1% formic acid, 80% acetonitrile in H2O) at a flow rate of 300 nL/min. MS analysis was done with a full MS scan from 350 to 1750 m/z in Orbitrap, followed by data-dependent MS/MS scan of the 20 most intense ions in ion trap. Microbiome samples corresponding to each individual were run on LC-MS/MS in a randomized order. Spectral data were collected as *.RAW files.
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!