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Easy spray pepmap rslc c18 column

Manufactured by Thermo Fisher Scientific
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The EASY-Spray PepMap RSLC C18 column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of peptides. It features a silica-based stationary phase with a C18 alkyl chain bonded to the surface, providing reversed-phase chromatographic separation. The column is designed for use in EASY-Spray LC-MS systems, providing consistent and reliable performance for peptide analysis applications.

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Lab products found in correlation

Ultimate 3000, by Thermo Fisher Scientific (6 mentions) Ultimate 3000 rslcnano system, by Thermo Fisher Scientific (6 mentions) Orbitrap fusion lumos, by Thermo Fisher Scientific (4 mentions) Q exactive high field mass spectrometer, by Thermo Fisher Scientific (4 mentions) Acclaim pepmap100, by Thermo Fisher Scientific (4 mentions) Acclaim pepmap c18, by Thermo Fisher Scientific (4 mentions) Easy spray ion source, by Thermo Fisher Scientific (3 mentions) Dionex ultimate 1000 system, by Thermo Fisher Scientific (3 mentions) Acclaim pepmap 100 c18, by Thermo Fisher Scientific (3 mentions) Dionex ultimate 3000, by Thermo Fisher Scientific (3 mentions) Ltq orbitrap velos pro, by Thermo Fisher Scientific (3 mentions) Q exactive hf mass spectrometer, by Thermo Fisher Scientific (2 mentions) C18 pepmap100, by Thermo Fisher Scientific (2 mentions) Ultimate 3000 rslcnano, by Thermo Fisher Scientific (2 mentions) Q exactive, by Thermo Fisher Scientific (1 mentions) Fusion lumos, by Thermo Fisher Scientific (1 mentions) Q exactive hybrid quadrupole orbitrap mass spectrometer, by Thermo Fisher Scientific (1 mentions) Acclaim pepmap100 c18 pre column, by Thermo Fisher Scientific (1 mentions) Fusion tribrid mass spectrometer, by Thermo Fisher Scientific (1 mentions) Ltq orbitrap velos pro mass spectrometer, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

EASY-Spray PepMap RSLC C18 75μm × 250mm column EASY-Spray PepMap RSLC C18 75 µm × 250 mm column EASY-Spray PepMap RSLC C18 PepMap RSLC EASY-Spray column EASY PepMapTM RSLC C18 column Easy-Spray RSLC C18 column Easy Spray PepMap C18 column Easy-Spray PepMap® RSLC Easy-Spray C18 PepMap Easy Spray PepMap column

24 protocols using easy spray pepmap rslc c18 column

1

Quantitative Proteomics by LC-MS/MS

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LC-MS/MS analysis was performed using a Dionex Ultimate 3000 nano-ultra high-pressure reverse-phase chromatography coupled on-line to a Q Exactive HF mass spectrometer (Thermo Scientific) as described previously (Fye et al., 2018 (link)). In brief, samples were separated on an EASY-Spray PepMap RSLC C18 column (500 mm × 75 μm, 2 μm particle size, Thermo Scientific) over a 60 min gradient of 2–35% acetonitrile in 5% dimethyl sulfoxide (DMSO), 0.1% formic acid at 250 nL/min. MS1 scans were acquired at a resolution of 60,000 at 200 m/z and the top 12 most abundant precursor ions were selected for high collision dissociation (HCD) fragmentation.
Pinto-Fernández A., Davis S., Schofield A.B., Scott H.C., Zhang P., Salah E., Mathea S., Charles P.D., Damianou A., Bond G., Fischer R, & Kessler B.M. (2019). Comprehensive Landscape of Active Deubiquitinating Enzymes Profiled by Advanced Chemoproteomics. Frontiers in Chemistry, 7, 592.
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2

Proteomics Analysis of Islet Samples

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A Dionex UltiMate 1000 system (Thermo Scientific) was coupled to an Orbitrap Fusion Lumos (Thermo Scientific) through an Easy-Spray ion source (Thermo Scientific). For the islet sample, nLC separation was carried out as follows, while other samples were analyzed as previously described21 (link). Peptide samples were loaded (15 μL/min, 1 min) onto a trap column (100 μm × 2 cm, 5 μm Acclaim PepMap 100 C18, 50 °C), eluted (0.2 μL/min) onto an Easy-Spray PepMap RSLC C18 column (2 μm, 50 cm × 75 μm ID, 50 °C, Thermo Scientific) and separated with the following gradient, all % Buffer B (0.1% formic acid in ACN): 0–110 min, 2%–22%; 110–120 min, 22%–35%; 120–130 min, 35–95%; 130–150 min, isocratic at 95%; 151–153 min, 95%–2%, 153–171 min, isocratic at 2%. Spray voltage was 1900V, ion transfer tube temperature was 275°C, and RF lens was 30%. MS scans were acquired in profile mode and MS/MS scans in centroid mode, for ions with charge states 2–7, with a cycle time of 3 sec. MS spectra were recorded from 375–1500 Da at 120K resolution (at m/z 200), and HCD MS/MS was triggered above a threshold of 2.0e4, with quadrupole isolation (0.7 Da) at 30K resolution, and collision energy (CE) of 30%. Dynamic exclusion was used (60 s), and monoisotopic precursor selection was on.
Wan X., Vomund A.N., Peterson O.J., Chervonsky A.V., Lichti C.F, & Unanue E.R. (2020). The MHCII peptidome of the pancreatic islet identifies key features of autoimmune peptides. Nature immunology, 21(4), 455-463.
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3

High-Sensitivity LC-MS/MS Proteome Analysis

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LC-MS/MS analysis was performed using a Dionex Ultimate 3000 nano-ultra high-pressure reverse-phase chromatography coupled on-line to a Q Exactive HF (GlyGly), Fusion Lumos (ISG15 interactome)39 or a Q Exactive (Matching proteome)40 mass spectrometer (Thermo Scientific) as described previously.38 (link) In brief, samples were separated on an EASY-Spray PepMap RSLC C18 column (500 mm × 75 μm, 2 μm particle size, Thermo Scientific) over a 60 min (120 min in the case of the matching proteome) gradient of 2–35% acetonitrile in 5% dimethyl sulfoxide (DMSO), 0.1% formic acid at 250 nL/min. MS1 scans were acquired at a resolution of 60,000 at 200 m/z and the top 12 most abundant precursor ions were selected for high collision dissociation (HCD) fragmentation.
Pinto-Fernandez A., Salio M., Partridge T., Chen J., Vere G., Greenwood H., Olie C.S., Damianou A., Scott H.C., Pegg H.J., Chiarenza A., Díaz-Saez L., Smith P., Gonzalez-Lopez C., Patel B., Anderton E., Jones N., Hammonds T.R., Huber K., Muschel R., Borrow P., Cerundolo V, & Kessler B.M. (2020). Deletion of the deISGylating enzyme USP18 enhances tumour cell antigenicity and radiosensitivity. British Journal of Cancer, 124(4), 817-830.
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4

Proteomics Analysis of Islet Samples

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A Dionex UltiMate 1000 system (Thermo Scientific) was coupled to an Orbitrap Fusion Lumos (Thermo Scientific) through an Easy-Spray ion source (Thermo Scientific). For the islet sample, nLC separation was carried out as follows, while other samples were analyzed as previously described21 (link). Peptide samples were loaded (15 μL/min, 1 min) onto a trap column (100 μm × 2 cm, 5 μm Acclaim PepMap 100 C18, 50 °C), eluted (0.2 μL/min) onto an Easy-Spray PepMap RSLC C18 column (2 μm, 50 cm × 75 μm ID, 50 °C, Thermo Scientific) and separated with the following gradient, all % Buffer B (0.1% formic acid in ACN): 0–110 min, 2%–22%; 110–120 min, 22%–35%; 120–130 min, 35–95%; 130–150 min, isocratic at 95%; 151–153 min, 95%–2%, 153–171 min, isocratic at 2%. Spray voltage was 1900V, ion transfer tube temperature was 275°C, and RF lens was 30%. MS scans were acquired in profile mode and MS/MS scans in centroid mode, for ions with charge states 2–7, with a cycle time of 3 sec. MS spectra were recorded from 375–1500 Da at 120K resolution (at m/z 200), and HCD MS/MS was triggered above a threshold of 2.0e4, with quadrupole isolation (0.7 Da) at 30K resolution, and collision energy (CE) of 30%. Dynamic exclusion was used (60 s), and monoisotopic precursor selection was on.
Wan X., Vomund A.N., Peterson O.J., Chervonsky A.V., Lichti C.F, & Unanue E.R. (2020). The MHCII peptidome of the pancreatic islet identifies key features of autoimmune peptides. Nature immunology, 21(4), 455-463.
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5

LC-MS/MS Analysis of Protein Samples

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Liquid chromatography tandem mass spectrometry (LC–MS/MS) analysis was performed using a Dionex Ultimate 3000 nano-ultra high pressure reversed-phase chromatography system coupled on-line to a Q Exactive High Field (HF) mass spectrometer (Thermo Scientific) as described previously [18 (link)]. In brief, samples were separated on an EASY-Spray PepMap RSLC C18 column (500 mm × 75 µm, 2 µm particle size; Thermo Fisher Scientific) over a 60 min gradient of 2–35% ACN in 5% dimethyl sulfoxide (DMSO), 0.1% FA, and the flow rate was ~ 250 nL/min. The mass spectrometer was operated in data-dependent analysis (DDA) mode for automated switching between MS (MS1) and MS/MS (MS2) acquisition. Full MS survey scans were acquired from 400–2000 m/z at a resolution of 60,000 at 200 m/z and the top 12 most abundant precursor ions were selected for high collision energy dissociation (HCD) fragmentation. The resolution of MS2 fragment ion detection was also set to 15,000.
Thorne A.M., Huang H., O‘Brien D.P., Eijken M., Krogstrup N.V., Norregaard R., Møller B., Ploeg R.J., Jespersen B, & Kessler B.M. (2020). Subclinical effects of remote ischaemic conditioning in human kidney transplants revealed by quantitative proteomics. Clinical Proteomics, 17, 39.
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6

Peptide Characterization by Orbitrap MS

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Each peptide sample was reconstituted in 10 µl 5% formic acid then diluted to 1% prior to MS analysis. Peptide samples were injected onto a C18 PepMap 100 (300 μm x 5 mm, Thermo Scientific) trap column with buffer A (0.1% formic acid in MS grade water) using an Ultimate 3000 RSLC nano UHPLC system. After a 5 min wash at 5 µl/min the sample was then eluted onto an EasySpray PepMap RSLC C18 column (75 µm x 50 cm, Thermo Scientific) into a LTQ Orbitrap Velos Pro via an EasySpray ion source. The peptides were eluted from the column using a flow rate of 300 nl/min and a linear gradient from 2% to 40% buffer B (0.08% formic acid in 80% MS grade CH3CN) over 124 min. The column temperature was set at 50 °C. The Orbitrap Velos Pro ms system was operated in data dependant acquisition mode using a Top 15 method with Lockmass = 445.120024. A MS1 survey scan with a range of 335–1800 m/z and a resolution of 60,000 was followed by 15 sequential MS2 scans at the normal scan rate using the LTQ Velos ion trap. The FTMS and ITMS AGC targets were set to 1e6 ions and 5e3 ions respectively. The FTMS and ITMS maximum fill times were set to 500 msec and 100 msec respectively. ITMS isolation width was set at 2 Da with a normalised collision energy of 35, a default charge state of 2, an Activation Q of 0.250 and Activation Time of 10 msec.
Gadd M.S., Testa A., Lucas X., Chan K.H., Chen W., Lamont D.J., Zengerle M, & Ciulli A. (2017). Structural basis of PROTAC cooperative recognition for selective protein degradation. Nature chemical biology, 13(5), 514-521.
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7

LC-MS Analysis of Biomolecules

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The LC–MS analysis was performed on a Q Exactive™ Hybrid Quadrupole-Orbitrap™ Mass Spectrometer (Thermo Scientific, San Jose, CA), coupled online to an Ultimate 3000 ultra-high performance liquid chromatography (UHPLC) instrument (Thermo Scientific, San Jose, CA). The UHPLC system was equipped with an Easy Spray Pepmap RSLC C18 column (2 µm particle, 100 Å pore size, and dimensions: 50 µm × 15 cm, Thermo Scientific, San Jose, CA). Before sample separation on the analytical column, the lyophilized sample was resuspended in 16 µL solution containing 2% (v/v) CH3CN and 0.1% (v/v) FA solution. Next, 5 µL sample was injected and loaded on an Acclaim Pepmap 100 C18 precolumn (3 µm particle size, 100 Å pore size, nanoviper, and dimensions: 75 µm × 2 cm, Thermo Scientific, San Jose, CA) at a flow rate of 5 μL/min. Sample separation was performed using a 95 min gradient. Mobile phase A consisted of 99.9% H2O and 0.1% (v/v) FA and mobile phase B of 19.92% H2O, 80% (w/v) CH3CN and 0.08% (v/v) FA. Mobile phase B increased from 4 to 10% in 5 min, 10–25% in 50 min, 25–45% in 18 min followed by a steep increase to 95% in 1 min. A flow rate of 300 nL/min was used. An inherent rinse step (10 min gradient, from 4–95% in 5 min) was applied after every 95 min separation gradient.
Zhang Z., Boonen K., Ferrari P., Schoofs L., Janssens E., van Noort V., Rolland F, & Geuten K. (2016). UV crosslinked mRNA-binding proteins captured from leaf mesophyll protoplasts. Plant Methods, 12, 42.
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8

LC-MS/MS Proteomic Analysis Protocol

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Liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis was performed in biological quadruplicate using a Dionex Ultimate 3000 nano-ultra high pressure reverse phase chromatography coupled on-line to a Q Exactive High Field (HF) mass spectrometer (Thermo Scientific) as described40 (link). Samples were separated on an EASY-Spray PepMap RSLC C18 column (500 mm × 75 µm, 2 µm particle size, Thermo Scientific) over a 60 min gradient of 2-35% acetonitrile in 5% DMSO, 0.1% formic acid at 250 nl/min. MS1 scans were acquired at a resolution of 60,000 at 200 m/z and the top 12 most abundant precursor ions were selected for HCD fragmentation.
Stangl A., Elliott P.R., Pinto-Fernandez A., Bonham S., Harrison L., Schaub A., Kutzner K., Keusekotten K., Pfluger P.T., El Oualid F., Kessler B.M., Komander D, & Krappmann D. (2019). Regulation of the endosomal SNX27-retromer by OTULIN. Nature Communications, 10, 4320.
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9

Nano-LC-MS/MS Proteomic Analysis

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Samples were analyzed at UNITECH OMICs (University of Milan, Milan, Italy) using a Dionex Ultimate 3,000 nano-LC system connected to an orbitrap Fusion Tribrid Mass Spectrometer (Thermo Fisher Scientific) equipped with a nanoelectrospray ion source operating in positive ion mode. The peptide mixtures were preconcentrated onto an Acclaim PepMap C18 5 µm, 100 Å, 100 µm ID × 2 cm (Thermo Fisher Scientific) and separated at 35°C on an EASY-Spray PepMap RSLC C18 column: 3 µm, 100 Å, 75 µm ID × 15 cm (Thermo Fisher Scientific). Each group was injected three times.
Elutions were run in gradient mode from 96% buffer A (0.1% aqueous formic acid) to 40% buffer B (0.1% aqueous formic acid/acetonitrile [2:8]). Run total length: 110 min, flow rate: 300 nl min−1. MS spectra were collected over an m/z range of 375—1,500 Da at a resolution of 120.000 in the data-dependent mode, cycle time 3 s between master scans. Fragmentation was induced by higher-energy collisional dissociation (HCD) with collision energy set at 35 eV.
Bonacina F., Moregola A., Svecla M., Coe D., Uboldi P., Fraire S., Beretta S., Beretta G., Pellegatta F., Catapano A.L., Marelli-Berg F.M, & Norata G.D. (2022). The low-density lipoprotein receptor–mTORC1 axis coordinates CD8+ T cell activation. The Journal of Cell Biology, 221(11), e202202011.
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10

Peptide Analysis by Nano-UPLC-MS/MS

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Peptides were analyzed using a Dionex Ultimate 3000 nano-ultra high pressure reversed-phase chromatography system coupled on-line to an Orbitrap Fusion Lumos mass spectrometer (Thermo Scientific). Samples were separated on an EASY-Spray PepMap RSLC C18 column (500 mm × 75 μm, 2 μm particle size; Thermo Scientific) over a 60 min gradient of 2 to 35% ACN in 5% DMSO, 0.1% FA, and at 250 nl/min. The column temperature was maintained at 50 °C with the aid of a column oven. The mass spectrometer was operated in positive polarity mode with a capillary temperature of 305 °C. DIA mode was utilized for automated switching between MS and MS/MS acquisition as described previously (28 (link)). Briefly, full scans (m/z 350–1650) were acquired in the Orbitrap with 120 k resolution and maximum injection time of 20 ms, followed by 40 DIA scan windows with variable widths (supplemental Table S1). MS/MS fragmentation was performed in the higher energy collisional dissociation cell with a collision energy set at 30%. MS2 scans were acquired in the Orbitrap between m/z 200 and 2000 at a resolution of 30 k. The minimum points-per-peak was enabled and set to 6 with a dynamic maximum injection. All data were acquired in profile mode.
O'Brien D.P., Jones H.B., Guenther F., Murphy E.J., England K.S., Vendrell I., Anderson M., Brennan P.E., Davis J.B., Pinto-Fernández A., Turnbull A.P, & Kessler B.M. (2023). Structural Premise of Selective Deubiquitinase USP30 Inhibition by Small-Molecule Benzosulfonamides. Molecular & Cellular Proteomics : MCP, 22(8), 100609.
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