How female falsewidowspiders use their 'spidey senses' to attract mates—study sfu CommsBio
, accounting for the different amounts of silk produced by these three groups of spiders. For each replicate, webs were extracted in 200 µL 0.05 M Sørensen bufferand analysed by Bioinformatics Solutions . After web samples were incubated for 20 min at 60 °C in 2× sample volumes of 10% SDS , they were sonicated for 20 min. Then, the supernatant was withdrawn, reduced with dithiothreitol , and alkylated with iodoacetamide . Alkylated samples were treated further with asolvent .
Eluates were analysed by HPLC-MS/MS in positive ion mode on a Thermo Scientific Orbitrap Fusion Lumos Tribrid mass spectrometer , equipped with a nanospray ionisation source and a Thermo Fisher Ultimate 3000 RSLCnano HPLC System . Peptide mixtures were loaded onto a PEPMAP100 Ctrap column at a constant flow of 30 μL/min and 60 °C isothermal. Peptides were eluted at a rate of 0.
MS data were acquired in data-dependent mode with a cycle time of 3 s. MS1 scan data were acquired with the Orbitrap mass analyser, using a mass range of 400–1600, with the resolution set to 120,000. The automatic gain control was set to 4e5, with a maximum ion injection time of 50 ms, and the radio frequency lens was set to 30%. Isolations for MS2 scans were run using a quadrupole mass analyser, with an isolation window of 0.7.
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