Peptide samples were loaded directly onto an in house packed reverse phase column using 5 µm, 200Å particles (magic C18, Michrom) and PicoTip Emmiters (New Objective) with an autosampler / nanoLC setup (2D nanoLC, Eksigent) at a flow rate of 1 µl/min. After loading the column was washed for 5 min at 1 µl/min at 99 %A (water with 0.2 % FA) 1 %B (acetonitrile with 0.2 % FA) followed by elution with a linear gradient from 1 % B to 35 % B at 400 nl/min in 60 min. Peptides eluting from the column were ionized in the positive ion mode and the 6 most abundant ions were fragmented in the PQD-mode30 (link) to allow for the detection of low mass range reporter ions. Briefly, the LTQ-Orbitrap was run in positive ion mode. Full scans were carried out with a scan range of 395 to 1200 m/z. Normalized collision energy of 35 was used to activate both the reporter ions and parent ions for fragmentation. Scans were carried out with an activation time of 30 ms. The isolation window was set to 1.0 m/z.
Picotip emmiters
Manufactured by New Objective
PicoTip Emitters are a type of lab equipment designed for the precise delivery of small volumes of liquids. They feature a tapered tip with a small orifice size, allowing for the controlled dispensing of micro- or nano-liter quantities. The core function of PicoTip Emitters is to enable accurate and reproducible sample introduction for various analytical techniques.
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Lab products found in correlation
2 protocols using picotip emmiters
1
Nano-LC-MS/MS Peptide Analysis Protocol
2
Nano-LC-MS/MS Peptide Analysis Protocol
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Peptide samples were loaded directly onto an in house packed reverse phase column using 5 µm, 200Å particles (magic C18, Michrom) and PicoTip Emmiters (New Objective) with an autosampler / nanoLC setup (2D nanoLC, Eksigent) at a flow rate of 1 µl/min. After loading the column was washed for 5 min at 1 µl/min at 99 %A (water with 0.2 % FA) 1 %B (acetonitrile with 0.2 % FA) followed by elution with a linear gradient from 1 % B to 35 % B at 400 nl/min in 60 min. Peptides eluting from the column were ionized in the positive ion mode and the 6 most abundant ions were fragmented in the PQD-mode30 (link) to allow for the detection of low mass range reporter ions. Briefly, the LTQ-Orbitrap was run in positive ion mode. Full scans were carried out with a scan range of 395 to 1200 m/z. Normalized collision energy of 35 was used to activate both the reporter ions and parent ions for fragmentation. Scans were carried out with an activation time of 30 ms. The isolation window was set to 1.0 m/z.
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