Venom and saliva samples were collected from 2 wild-caught adult male Hispaniolan solenodons. We used reduced SDS-PAGE gel electrophoresis for initial visualization of venom and saliva proteins (10 µg). Shotgun proteomics was performed by digesting 5 µg of each sample with trypsin, before analysis by LC-MS/MS. Decomplexed bottom-up proteomics were performed as previously described (27 (link)). Samples (1 mg) were separated via reverse-phase HPLC, then reduced and analyzed by SDS-PAGE, and protein bands subjected to in-gel tryptic digestion and analyzed by LC-MS/MS using an Orbitrap XL (Agilent, Germany). For top-down proteomics we used 0.2 mg of venom and saliva for reduced and nonreduced HPLC high-resolution (HR) MS/MS measurements. Top-down LC-electrospray ionization-HR-MS experiments were performed on an LTQ Orbitrap XL (Agilent) in data-dependent acquisition mode. For all proteomic experiments, resulting MS2 spectra (57 ) were matched against translations of the protein-encoding genes predicted from the genome. Full details are displayed in SI Appendix, File S1 .
Orbitrap xl
Manufactured by Agilent Technologies
Sourced in Germany
The Orbitrap XL is a high-resolution mass spectrometer designed for accurate mass measurements and in-depth analysis of complex samples. It utilizes Orbitrap mass analyzer technology to provide high mass resolution, mass accuracy, and sensitivity for a wide range of applications.
Automatically generated - may contain errors
2 protocols using orbitrap xl
1
Proteomic Analysis of Hispaniolan Solenodons
2
Quantitative Mass Spectrometry Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LC/MS was performed on a Thermo Scientific Orbitrap XL mass spectrometer linked to an Agilent1200 nano-high-performance liquid chromatography. Peptides were resolved in 4% acetonitrile and 0.5% formic acid and loaded on a C18 reversed-phase precolumn (Zorbax 300SB-C18; Agilent Technologies, Palo Alto, CA) followed by separation on an in-house packed 3 mm Reprosil C18 resin (Dr. Maisch GmbH, Ammerbuch, Germany) picotip emitter tip (diameter 75 mm, length 10 cm; New Objectives, Woburn, MA) using a gradient from 5% acetonitrile, 0.1% formic acid to 50% acetonitrile, 0.1% formic acid for 90 minutes with a flow rate of 200 nL/min. MS data were recorded by data-dependent Top10 acquisition selecting the 10 most abundant precursor ions in positive mode for fragmentation using dynamic exclusion of 3 minutes. Only highly charged ions (21) were selected for MS/MS scans in the linear ion trap by CID at 35% collision energy. Lock mass option for m/z 445.120,025 was enabled to ensure high mass accuracy during many following runs. 57
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!