Acclaim pepmap rslc c18 reverse phase column

Manufactured by Thermo Fisher Scientific

The Acclaim PepMap RSLC C18 reverse phase column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of peptides and proteins. The column features a C18 stationary phase and is optimized for rapid and efficient separations.

Automatically generated - may contain errors

Lab products found in correlation

Easy nlc 1000 liquid chromatography system, by Thermo Fisher Scientific (1 mentions) Orbitrap fusion tribrid, by Thermo Fisher Scientific (1 mentions) Nupage bis tris gel, by Thermo Fisher Scientific (1 mentions) Lds loading buffer, by Thermo Fisher Scientific (1 mentions) Orbitrap fusion mass spectrometer, by Thermo Fisher Scientific (1 mentions) Ultimate 3000 lc system, by Thermo Fisher Scientific (1 mentions) Trypsin, by Roche (1 mentions)

Close spelling variants of this product

Acclaim PepMap RSLC C18 column Acclaim PepMap RSLC C18 Acclaim PepMap RSLC column Acclaim RSLC C18 column PepMap RSLC C18 column Acclaim PepMap C18 column Acclaim PepMap RSLC PepMap RSLC C18 Acclaim PepMap C18 PepMap RSLC column

2 protocols using acclaim pepmap rslc c18 reverse phase column

Proteolytic Peptide Separation and Identification

Check if the same lab product or an alternative is used in the 5 most similar protocols

To analyze the digests from the CL experiments, a Thermo Scientific EASY-nLC 1000 liquid chromatography system (Thermo Scientific, Tewksbury, MA) with an Acclaim PepMap RSLC C18 reverse phase column (75 μM × 15 cm, 2 μm particle size) from Thermo Scientific (Tewksbury, MA) was used. To achieve efficient separation of the proteolytic peptides, a gradient was used in which the %B (0.1% formic acid in acetonitrile) was increased from 0% to 40% over 45 min. The column was then flushed by increasing to 95% B over 15 min. The column was then cleaned at 95% B for another 20 min. A flow rate of 300 nL/min was used throughout the run.
LC-MS and LC-MS/MS analyses of protein proteolytic fragments were conducted on a Thermo Orbitrap Fusion Tribrid (Tewksbury, MA) mass spectrometer. The electrospray ionization source was typically operated at a needle voltage of 2100 V, and the ion transfer tube temperature was set to 300 °C. Tandem mass spectra were collected using CID with a normalized collision energy of 35%. Due to the large number of detectable peaks, an exclusion limit of 60 s was applied after five spectra had been collected for any given peak. The resolution of the Orbitrap was set to 60000. Peptides were analyzed and identified using the Thermo Proteome Discoverer 2.2 software or via manual analysis. Search details are provided in the SI.

Zhao B., Zhuang J., Xu M., Liu T., Limpikirati P., Thayumanavan S, & Vachet R.W. (2020). Covalent Labeling with an α, β Unsaturated Carbonyl Scaffold for Studying Protein Structure and Interactions by Mass Spectrometry. Analytical chemistry, 92(9), 6637-6644.

+ Open protocol

+ Expand

Membrane Proteome Isolation and Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

IM and OM samples were carefully dissolved in 100 μL 1X LDS loading buffer (Invitrogen) before loading on a precast Tris-Bis NuPAGE gel (Invitrogen) using 1X MOPS running solution (Invitrogen). SDS-polyacrylamide gel electrophoresis was run for approximately 5 min to purify polypeptides in the polyacrylamide gel by removing contaminants. Polyacrylamide gel bands containing the membrane proteome were excised and digested by trypsin (Roche) proteolysis. The resulting tryptic peptides were extracted using formic acid-acetonitrile (5%:25%, v/v) before resuspension in acetonitrile-trifluoroacetate (2.5%:0.05%, v/v). Tryptic peptides were separated by nano-liquid chromatography (nanoLC) using an Ultimate 3000 LC system with an Acclaim PepMap RSLC C18 reverse phase column (ThermoFisher) at the Proteomics Research Technology Platform (PRTP) at the University of Warwick. MS/MS spectra were collected using an Orbitrap Fusion mass spectrometer (ThermoFisher) in electrospray ionization (ESI) mode. Survey scans of peptides from m/z 350 to 1500 were collected for each sample in a 1.5-hr LC-MS run. This resulted in 12 mass spectra (3 biological replicates of IM and OM of WT and the olsA mutant) with a total of ~ 7.5 G of MS/MS data.

Stirrup R., Mausz M.A., Silvano E., Murphy A., Guillonneau R., Quareshy M., Rihtman B., Ferretjans M.A., He R., Todd J.D., Chen F., Scanlan D.J, & Chen Y. (2022). Aminolipids elicit functional trade-offs between competitiveness and bacteriophage attachment in Ruegeria pomeroyi. The ISME Journal, 17(3), 315-325.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!