Cannabivarin and Tetrahydrocannabivarin Modulate Nociception via Vanilloid Channels and Cannabinoid-Like Receptors in Caenorhabditis elegans dataset2
Description
Dataset 2 for the project Cannabivarin and Tetrahydrocannabivarin Modulate Nociception via Vanilloid Channels and Cannabinoid-Like Receptors in Caenorhabditis elegans
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Steps to reproduce
Proteomics analysis Online chromatographic separation was performed using a Thermo Scientific Vanquish Neo UHPLC system (San Jose, CA, USA) configured in trap-and-elute mode. The setup included a Thermo Scientific PepMap Neo 5 μm C18 300 μm × 5 mm trap cartridge and a Thermo Scientific PepMap Neo C18 2 μm × 75 μm × 500 mm nano column. A 2 μL sample (∼2 μg of digested proteins) was loaded onto the trap column and desalted with solvent A [0.1% formic acid in water] at a 20 μL/min flow rate for 0.5 minutes. Peptide elution was carried out using a linear gradient of 5% to 35% solvent B [80% acetonitrile, 20% water, 0.1% formic acid] over 135 minutes, followed by a 2-step column wash at 50% solvent B for 10 minutes and 80% for 10 minutes. The flow rate was maintained at 200 nL/min. Afterwards, the column was equilibrated to the initial solvent composition (5% solvent B) by running 10 column volumes. Data acquisition was conducted on a Thermo Scientific Q Exactive Plus Orbitrap Mass Spectrometer (San Jose, CA, USA) equipped with a Nanospray Flex ion source. Data collection was performed in positive ion mode with a nanospray voltage of 2.2 kV, and the ion transfer tube was maintained at 220°C. The mass spectrometer was operated in data-dependent acquisition (DDA) mode, utilizing a TOP-12 acquisition strategy. A high-resolution MS1 survey scan (m/z 375– 1200) was acquired at a resolution of 70,000 (FWHM), with an automatic gain control (AGC) target value of 1 × 10 and a maximum injection time of 100 ms. The 12 most intense precursor ions exceeding an intensity threshold of 1 × 10 were selected for fragmentation via higher-energy collisional dissociation (HCD) at a normalized collision energy (NCE) of 27. MS2 spectra were acquired at a resolution of 17,500 (FWHM) using an AGC target value of 2 × 10 and a maximum injection time of 100 ms.
Institutions
- Universite de Montreal Faculte de medecine veterinaire