Fused silica emitter

Manufactured by New Objective

Sourced in United States

The Fused-silica emitter is a laboratory instrument designed to produce a focused stream of ions or charged particles. It features a fused-silica capillary that generates a stable electrospray for various analytical applications. The core function of this product is to facilitate the introduction of samples into mass spectrometry or other analytical systems.

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Lab products found in correlation

Spelling variants of this product

20 cm fused silica emitter (5 citations) 30 μm fused silica emitter (2 citations) 23-cm fused silica emitter (2 citations) Fused silica emitter tip (2 citations)

6 protocols using fused silica emitter

Nanoflow LC-MS Peptide Separation

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Corresponding to
DNA amounts ranging from 1–8 μg (cell samples) or 15–50
μg (ctDNA samples), 0.5, 2, or 5 μL of hydrolyzed, SPE-purified,
and reconstituted sample was injected onto a nanoACQUITY UPLC (Waters
Co., Milford, MA) system equipped with a 5 μL injection loop.
Separation was performed with a capillary column (75 μm ID,
10 cm length, 15 μm orifice) created by hand packing a commercially
available fused-silica emitter (New Objective, Woburn MA) with 5 μm
Luna C18 bonded separation media (Phenomenex, Torrance, CA). The flow
rate was 1000 nL/min for 5.5 min, then decreased to 300 nL/min with
a 50 min linear gradient from 2–50% CH₃CN in 5 mM
NH₄OAc aqueous buffer (pH 5.5), an increase to 98% CH₃CN in 3 min, with a 2 min hold and a 5 min re-equilibration
at 1000 nL/min at 2% CH₃CN. The injection valve was switched
at 5.5 min to remove the sample loop from the flow path during the
gradient.

Stornetta A., Villalta P.W., Gossner F., Wilson W.R., Balbo S, & Sturla S.J. (2017). DNA Adduct Profiles Predict in Vitro Cell Viability after Treatment with the Experimental Anticancer Prodrug PR104A. Chemical Research in Toxicology, 30(3), 830-839.

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High-Resolution Nano-LC-MS Peptide Separation

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Chromatographic separation was performed using a Dionex UltiMate 3000 RSLCnano UPLC system coupled to the nanospray ion source containing a capillary column (75 μm ID, 15 cm length, 10 μm orifice) created by hand packing a commercially available fused-silica emitter (New Objective, Woburn, MA) with XBridge BEH C18 (5 μm particle size, 130 Å pore size) stationary phase (Waters, Milford, MA). The LC solvents were: (A) 0.05% CH₃COOH in H₂O and (B) CH₃CN. Three μL injections were performed and the initial flow rate and composition of 5% B and 600nl/min was held for 10 min to push sample out of the 5 μL autosampler loop, followed by switching the injection valve from the inject to load position to take the loop out of the flow path, and a decrease of the flow rate to 300 nL/min over the course of 1 min. This was followed by a linear gradient of the B composition over 6.5 min to 61.5% B at which point the flow rate was decreased to 150 nL/min and held at 61.5% B for 2 min. The column was re-equilibrated between injections at the initial conditions for 9 min. To assure maximum capture of the 3 μL injection from the 3 μL sample volume, the sampler sample height was set to 0.0 mm in the Xcalibur method and the sample container height offset was set to 0.0 mm in the DCMSLink Chromeleon Xpress UPLC control software.

Villalta P.W., Hochalter J.B, & Hecht S.S. (2017). Ultra-sensitive High Resolution Mass Spectrometric Analysis of a DNA Adduct of the Carcinogen Benzo[a]pyrene in Human Lung. Analytical chemistry, 89(23), 12735-12742.

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N-Glycan Profiling by Orbitrap MS

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The samples were sodiated by 1 mM NaOH in 80% MeOH then directly infused onto a 30 µm fused silica emitter (New Objective). The detection of N-linked glycans was performed on the Q Exactive™ Plus Orbitrap Mass Spectrometer (Thermo Scientific) furnished with a Nanospray Flex Ion Source for direct infusion at 0.5 µL/min flow rate. The full MS spectra were obtained for relative quantitative analysis with 30 s data acquisition time at 600 to 2000 Da mass range.

Yue X., Tiwari N., Zhu L., Ngo H.D., Lim J.M., Gim B., Jing S., Wang Y., Qian Y, & Lee I. (2021). Tankyrase-1-mediated degradation of Golgin45 regulates glycosyltransferase trafficking and protein glycosylation in Rab2-GTP-dependent manner. Communications Biology, 4, 1370.

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Separation and Analysis of ctDNA by nanoLC-MS

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2.5 μg (ctDNA samples) or 1.5–4.6 μg (HT-29 samples) of DNA was injected onto a NanoLC-Ultra 2D HPLC (Eksigent, Dublin, CA) system equipped with a 5 μL injection loop. Separation was performed with a capillary column (75 μm ID, 10 cm length, 15 μm orifice) created by hand packing a commercially available fused-silica emitter (New Objective, Woburn MA) with 5 μm Luna C18 bonded separation media (Phenomenex, Torrance, CA). The flow rate was 1000 nL/min for 5 min, then decreased to 300 nL/min with a 50 min linear gradient from 2 to 98% CH₃CN in 5 mM NH₄OAc aqueous buffer (pH 6.8) with a 5 min hold and a 5 min re-equilibration at 1000 nL/min 98:2 buffer/CH₃CN. The injection valve was switched at 6 min to remove the sample loop from the flow path during the gradient. Samples were analyzed by nanoelectrospray using an LTQ Orbitrap Velos instrument (Thermo Scientific, Waltham, MA). The nanoelectrospray source voltage was 2.0 kV, and the capillary temperature was 350 °C. The ion focusing and transfer elements of the instrument were adjusted for maximum signal intensity by using the automated instrument tuning feature while monitoring the background ion signal of m/z 371.1 (decamethylcyclopentasiloxane) to create the tune file used for data analysis. This resulted in an S-Lens RF level setting of 49%.

Stornetta A., Villalta P.W., Hecht S.S., Sturla S.J, & Balbo S. (2015). Screening for DNA Alkylation Mono and Crosslinked Adducts with a Comprehensive LC-MS3 Adductomic Approach. Analytical chemistry, 87(23), 11706-11713.

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LC-MS/MS Analysis of DNA Adducts

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Samples were dissolved in 20 μL of H₂O for LC-NSI-HRMS/MS analysis on an Orbitrap Fusion Lumos instrument (Thermo Scientific, San Jose, CA) interfaced with a UPLC system (Ultimate 3000 RSLCnano UPLC, Thermo Scientific, Waltham, MA) using nanoelectrospray ionization. The UPLC was equipped with a 5 μL loop and the separation was carried out using a capillary column (75 μm ID, 20 cm length, 15 μm orifice) prepared by hand packing a commercially available fused-silica emitter (New Objective, Woburn MA) with Luna C₁₈ 5µ bonded separation media (Phenomenex, Torrance, CA). Parameters and conditions used for this analysis were previously reported.^{31 (link)} Extracted ion chromatograms were generated as follows: m/z 324 → 208.0829 for ɣ-OH-Acr-dGuo (1) and m/z 339 → 218.0849 for [¹³C₁₀¹⁵N₅]ɣ-OH-Acr-dGuo; m/z 338 → 222.0986 for (6S,8S)ɣ-OH-Cro-dGuo (2) and (6R,8R)ɣ-OH-Cro-dGuo (3) and m/z 343 → 227.0832 for [¹⁵N₅](6S,8S;6R,8R)ɣ-OH-Cro-dGuo; m/z 276 → 160.0618 for 1,N⁶-etheno-dAdo (4) and m/z 281 → 160.0618 for [¹³C₅]1,N⁶-etheno-dAdo; and m/z 284 → m/z 168.0511 for 8-oxo-dGuo (5) and m/z 287 → 171.0485 for [¹³C¹⁵N₂]8-oxo-dGuo.

Davidson J.J., Ozçelik T., Hamacher C., Willems P.J., Francke U, & Kilimann M.W. (1992). cDNA cloning of a liver isoform of the phosphorylase kinase alpha subunit and mapping of the gene to Xp22.2-p22.1, the region of human X-linked liver glycogenosis.. Proceedings of the National Academy of Sciences of the United States of America, 89(6), 2096-2100.

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Orbitrap-Based Proteomics Pipeline

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The LC/MS/MS runs were conducted on Orbitrap Exactive and Orbitrap Fusion mass spectrometers (Thermo Fisher Scientific, Bremen, Germany) connected to an Acquity nanoUPLC (Waters, Eschborn, Germany). Two μL of each sample were loaded onto a reversed-phase (RP) trapping column (Symmetry C18 Trap Column; 100 Å, 5 μm, 180 μm × 20 mm) and washed with 1% buffer B for 5 min. The peptides were eluted onto a RP capillary column (nanoAcquity Peptide BEH analytical column; 130 Å, 1.7 µm, 75 μm × 200 mm, buffer A: 0.1% formic acid in H₂O; buffer B: 0.1% formic acid in acetonitrile) and separated under the following conditions: 10 min at 3% solvent B followed by a gradient from 3% to 35% buffer B for 35 min (flow: 250 nL/min). Eluting peptides were ionized by ESI in the positive mode using a fused silica emitter (I.D. 10 μm, New Objective, Woburn, MA, USA) at a capillary voltage of 1,800 V.

Albrecht E., Schering L., Buck F., Vlach K., Schober H.C., Drevon C.A, & Maak S. (2020). Irisin: Still chasing shadows. Molecular Metabolism, 34, 124-135.

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