The peptides were synthesized by the F-moc strategy in a synthesizer machine (Multipep1; Intavis Bioanalytical Instruments, Köelh, Germany) using the Tentagel® resin as described elsewhere [64 (link)]. Peptides were purified by an HPLC coupled to mass spectrometry. The chromatographic column used was the XBridge BEH C18 from Waters, with a particle size of 2.7 μM and dimensions of 5 cm × 4.6 mm. The mobile phase was comprised of the following eluents: 0.05% formic acid in H2O (18 MΩ × cm) and 0.05% formic acid in acetonitrile. The equipment used was a Waters Autopurification System with a Waters 2545 binary gradient pump, a Waters 2998 diode array detector, and a Waters SQ Detector 2 electrospray ionization source mass detector (IBMP 002063). The diode array detector performed monitoring from 200–300 nm, and the mass detector served monitoring from m/z 450–1450 in positive mode. The injection volume used was 20 µL of a sample with an estimated 1 mg/mL concentration. The chromatography column was kept at room temperature.
2545 binary gradient pump
Manufactured by Waters Corporation
The 2545 binary gradient pump is a high-performance liquid chromatography (HPLC) pump designed for precise and reliable solvent delivery. It features a binary gradient configuration, allowing for the efficient mixing and delivery of two different solvents. The pump is capable of operating at a maximum pressure of 6,000 psi and flow rates ranging from 0.001 to 10.000 mL/min. The 2545 binary gradient pump is a key component in HPLC systems, providing the necessary solvent flow for separation and analysis of various samples.
Automatically generated - may contain errors
Lab products found in correlation
Xbridge beh c18, by Waters Corporation (1 mentions)
Autopurification system, by Waters Corporation (1 mentions)
Sq detector 2, by Waters Corporation (1 mentions)
2489 uv vis detector, by Waters Corporation (1 mentions)
Luna 5 μm silica column, by Phenomenex (1 mentions)
High performance liquid chromatography (hplc), by Waters Corporation (1 mentions)
2 protocols using 2545 binary gradient pump
1
Peptide Synthesis and Purification
2
Photostability Characterization of Fluorescent Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For WinterGreen-WAY
characterization, the compound was taken up in DMSO as a 1.5 mg/mL
solution and sterile-filtered to remove any aggregates. 100 μL
aliquots were irradiated with a 4500 mW/cm2 spot Hg-arc
light source (Hamamatsu LC8) for various time points. Aliquots were
then purified on a Waters HPLC, with a Waters 2545 binary gradient
pump equipped with Waters 2489 UVVis Detector. The column was a Phenomenex
C18 column (4.6 × 50 mm) run with a gradient of 5–95%
acetonitrile in water for 20 min. Percent WinterGreen-WAY remaining
was calculated from the integrated peak area.
For WinterRed-NDMC
characterization, compound was taken up in ethyl acetate as a 1.5
mg/mL solution and sterile-filtered to remove any aggregates. Irradiation
procedure and HPLC instrument were the same as above. The column was
a Phenomenex Luna 5 μm silica column (4.6 × 50 mm) run
with a gradient of 5–95% ethyl acetate in hexanes for 20 min.
Percent WinterRed-NDMC remaining was calculated from the integrated
peak area.
characterization, the compound was taken up in DMSO as a 1.5 mg/mL
solution and sterile-filtered to remove any aggregates. 100 μL
aliquots were irradiated with a 4500 mW/cm2 spot Hg-arc
light source (Hamamatsu LC8) for various time points. Aliquots were
then purified on a Waters HPLC, with a Waters 2545 binary gradient
pump equipped with Waters 2489 UVVis Detector. The column was a Phenomenex
C18 column (4.6 × 50 mm) run with a gradient of 5–95%
acetonitrile in water for 20 min. Percent WinterGreen-WAY remaining
was calculated from the integrated peak area.
For WinterRed-NDMC
characterization, compound was taken up in ethyl acetate as a 1.5
mg/mL solution and sterile-filtered to remove any aggregates. Irradiation
procedure and HPLC instrument were the same as above. The column was
a Phenomenex Luna 5 μm silica column (4.6 × 50 mm) run
with a gradient of 5–95% ethyl acetate in hexanes for 20 min.
Percent WinterRed-NDMC remaining was calculated from the integrated
peak area.
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