To identify expressed proteins, gel spots were excised and digested with trypsin. Tryptic peptides were subjected to nLC-MS/MS analysis using Thermo LTQ XL linear ion trap MS interfaced with nano LC system. 1 μL sample was injected through an autosampler into the nLC system at the flow rate of 300 nL/min. The peptides were eluted onto a 75 μm I.D ×15 cm Pep-Map 100 C-18 nanocolumn. The column was equilibrated with 96.8% A (0.1% F.A) and 3.2% B (98% ACN, 2% water, 0.1% F.A). Peptide separation was achieved with multi-step gradient from 3.2% to 80% solution B over 70 mins. Silica tip 10 +/− 1 μm emitter was used to apply the peptides on to LTQ MS through nanoelectrospray (NSI). The capillary voltage was set at 29.95 V and the capillary temperature was maintained at 270°C. Xcalibur software version 2.0.7SP1 was used to produce RAW files and Proteome Discoverer 1.2 to convert RAW files into MGF files (see Supplementary Material available online at http://dx.doi.org/10.1155/2014/532953 ). Protein identification was performed using Mascot search engine against latest version of Swiss-PROT. For Mascot search, following parameters were used: peptide mass tolerance ± 1.5 Da, MS/MS tolerance ± 0.5 Da, carbamidomethylation of Cys, and Met Oxidation as fixed and variable modifications, respectively.
Ltq xl linear ion trap ms
Manufactured by Thermo Fisher Scientific
Sourced in China
The LTQ XL™ Linear Ion Trap MS is a high-performance mass spectrometer designed for a variety of analytical applications. It utilizes a linear ion trap technology to efficiently capture and analyze ions, providing high sensitivity and resolution. The instrument is capable of conducting various mass spectrometry techniques, including MS and MS/MS analysis.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using ltq xl linear ion trap ms
1
Proteomic Identification of Expressed Proteins
2
Synthesis and Characterization of BO- and CM-Modified RNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BO-modified RNA and CM-modified RNA were prepared by reacting BO-Br and CM-Br with RNA containing PS-2′-OMe modifications, respectively. To a 20 μL solution of 50 μM RNA in 50 mM sodium phosphate buffer at pH 6.0 was added 20 μL 25 mM BO-Br or CM-Br in DMF. The solution was kept on a roller at room temperature for 48 h. The resulting solution was purified by Amicon-10K ultrafilters using water to remove excess BO-Br or CM-Br and desalted for 8 times. The concentration of the modified RNA solution was quantified by the standard UV260 method, and the stock solution was diluted for desired concentrations. Denatured PAGE and ESI-MS were used to characterize the modified RNA products. ESI-MS measurement was either provided by Hippobio (Zhejiang, China), Sango Biotech (Shanghai, China) or performed on a Thermo Scientific LTQ XL™ Linear Ion Trap MS at Tsinghua University. Upon storage at −20 °C, BO-RNA and CM-RNA were found stable in stock solution for at least 6 months.
3
Sulfur-Fluoride Exchange-Modified Aptamers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SF-modified aptamers, including mSuFEx-aptamers, was prepared by reacting Br-SF with PScontaining aptamers shown in Table S1. To a 50 μL solution of 100 μM PS-containing aptamers in 100 mM sodium phosphate buffer at pH 6.0 was added 50 μL 30 mM Br-SF in CH3CN. The solution was kept at 37 o C for 1 day. The resulting solution was combined with 200 μL water and extracted by 200 μL butanol twice. The bottom (water) phase was purified by Amicon-10K ultrafilters using water for 6 times. The concentration of the final SF-modified aptamer solution was quantified by the standard UV260 method, and the stock solution was diluted to desired concentrations for other experiments.
Denatured polyacrylamide gel electrophoresis (PAGE) and electrospray ionization (ESI)-ion trap mass spectrometry (MS) were used for characterizing the SF-modified aptamer products. The ESI-MS measurement was provided by Genscript (Beijing, China) or performed on a Thermo Scientific LTQ XL™ Linear Ion Trap MS at Tsinghua University. Upon storage at -20 o C, the BSF-modified aptamer products were found stable in stock solution for at least 1 months.
Denatured polyacrylamide gel electrophoresis (PAGE) and electrospray ionization (ESI)-ion trap mass spectrometry (MS) were used for characterizing the SF-modified aptamer products. The ESI-MS measurement was provided by Genscript (Beijing, China) or performed on a Thermo Scientific LTQ XL™ Linear Ion Trap MS at Tsinghua University. Upon storage at -20 o C, the BSF-modified aptamer products were found stable in stock solution for at least 1 months.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!