The method was developed using a Thermo Fisher UHPLC Dionex Ultimate 3000 (Germering, Germany) coupled to photodiode array detector model 3000 RS (Germering, Germany) equipped with ISO-3100SD pump, WPS 3000 SL autosampler and a TCC-3000 SD column thermostat. The separation was achieved on a Prontosil Kroma plus® C18 column (150 × 4.6 mm; 5 μm particle size). The software was chromeleon 6.8 (Germering, Germany). Deionized water was obtained from Milli-Q ultrapure water purification system (Thermo Scientific Barnstead Smart2Pure 3 UV, Hungary). Degassing of the mobile phase was carried out using an ultra sonicator (Elmasonic S 60 (H), Germany). The pH of the mobile phase was adjusted using Jenway pH-meter 3310 (Dunmow, Essex, United Kingdom). Samples were centrifuged using (Centurion K241R-United Kingdom). The spiked plasma samples were mixed using a vortex device (Velp scientifica, Europe). Rotatory Vacuum Concentrator (DVP-TYRO 12, Germany) for solvents evaporation equipped with vacuum pump, a solvent trap (CHRIST CT 02–50, Germany) and a rotor (CHRIST RVC 2–18 CDplus, Germany).
Ct 02 50
Manufactured by Martin Christ
Sourced in Germany
The CT 02–50 is a freeze-dryer (lyophilizer) manufactured by Martin Christ. It is used for the gentle drying of various materials, such as pharmaceuticals, food, or biological samples, through the process of sublimation. The CT 02–50 has a chamber volume of 50 liters.
Automatically generated - may contain errors
3 protocols using ct 02 50
1
UHPLC-PDA Analysis of Plasma Samples
2
Peptide Digestion and Desalting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
Centrifuge at 1000×g for 5 min at 20°C–25°C to pellet the beads and then carefully transfer the supernatant with digested peptides into a clean 1.5 mL-tube.
Resuspend the beads in additional 300 μL ddH2O, vortex briefly, pellet the beads by centrifugation for 5 min at 1000×g at 20°C–25°C, and then transfer the supernatant to the clean 1.5 mL-tube with the transferred digested peptides. Repeat this step 3 times.
Add 2 μL of 100% Trifluoroacetic acid (TFA) to acidify the diluted digest.
The digested peptides were desalted with a C-18 column. Add the next solution after each solution completely flows through by gravity. Add 1 mL of 100% methanol to the C-18 column, allow it to flow through and discard the effluent. Add 2 mL of ddH2O to the cartridge, allow it to flow through and discard the effluent. Add the acidified digest to the C-18 column, allow it to flow through by gravity, and then add the effluent back to the C-18 column. Add 1 mL of 5% methanol to the C-18 column and discard the effluent. Add 200 μL of 80% methanol to the C-18 column and collect the effluent with a new 1.5 mL tube. Repeat one time. Dry the desalted peptide solution with a vacuum centrifugal concentrator (Christ, CT02-50), centrifuging at 1300 rpm for 2 h at −60°C. Keep them at −20°C until ready for MS analysis.
3
NMR Characterization of Agl24 Product
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HPLC-purified Agl24 products (0.5mg-2mg) were dried using a Christ RVC 2-25 speed vacuum fitted with a Christ CT 02-50 cold-trap to remove excess acetonitrile, then freeze dried (Alpha 1-2 LDplus, Christ) to remove residual water. Products were subsequently dissolved in 600 µl of D2O and NMR spectra were recorded at 293K. The spectra were acquired on a Bruker AVANCE III HD 500 MHz NMR Spectrometer equipped with a 5 mm QCPI cryoprobe. For 1D TOCSY experiments, H1' was irradiated at 5.35 ppm, H2' was irradiated at 3.90 ppm, and H1" was irradiated at 4.49 ppm (Fig S8 ). A combination of 1 H 1 H COSY, 1D TOCSY, and 1 H 13 C HSQC experiments were used to fully assign the 1 H and 13 C signals for the Agl24 reaction product. Full 1 H and 13 C chemical shift assignments can be found in Table 1 and are recorded with respect to the residual HDO signal at 4.7 ppm.
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!