All peptides used in this study were synthesized using a solid-phase Fmoc method at Beijing Scilight Biotechnology LLC (Beijing, China), purified by HPLC (purity >95%), and characterized by MALDI−TOF MS. The sources of HIV-1SF33 and TZM-bl, as well as the methods used for their preservation and passage have been described previously [20] (link), [41] (link). The clinical isolates of HIV-1 subtypes B′, BC, and AE were obtained from treatment-naive patients and stored at our laboratory in Chinese Center for Disease Control and Prevention [46] (link), [47] (link). These viruses represent the predominant strains circulating in China from 2005 to 2008. Sprague-Dawley rats (male, 7 weeks old, and 150–180 g) were purchased from Beijing HFK Bioscience (Beijing, China) and fed for 2 days before being used for pharmacokinetic analysis. mPEG-MAL (2 and 5 kDa) was purchased from Beijing Kaizheng Biotech Development (Beijing, China). All of the other chemicals used in this study were purchased as the analytical grade from various commercial sources. HPLC analyses were performed on an Agilent 1200 system (Agilent, USA) equipped with a UV detector. MALDI-TOF mass spectra were recorded on a Bruker BIFLEX III spectrometer (Bruker Daltonics, Germany). All experiments with the viruses were performed in an approved biosafety level (BSL-3) containment laboratory.
Biflex 3 spectrometer
Manufactured by Bruker
Sourced in Germany
The BIFLEX III is a time-of-flight mass spectrometer from Bruker. It is designed to provide high-resolution mass analysis of a variety of sample types. The core function of the BIFLEX III is to accurately measure the mass-to-charge ratio of ions generated from the sample.
Automatically generated - may contain errors
Lab products found in correlation
U 3010 spectrophotometer, by Hitachi (2 mentions)
Sprague dawley rats, by Beijing Huafukang Biotechnology (1 mentions)
Agilent 1200 system, by Agilent Technologies (1 mentions)
F 4500 fluorescence spectrophotometer, by Hitachi (1 mentions)
Absolute pl quantum yield spectrometer c11347, by Hamamatsu Photonics (1 mentions)
H2so4, by Merck Group (1 mentions)
C2h5oh, by Merck Group (1 mentions)
Ch2cl2, by Merck Group (1 mentions)
Ch3cn, by Merck Group (1 mentions)
D ribose, by Merck Group (1 mentions)
F 7000 spectrophotometer, by Hitachi (1 mentions)
Avance 400 spectrometer, by Bruker (1 mentions)
Av 400 spectrometer, by Bruker (1 mentions)
Jsm 6301f, by JEOL (1 mentions)
Jms 700, by JEOL (1 mentions)
Advance drx 300, by Bruker (1 mentions)
Dm2500p microscope, by Leica (1 mentions)
7 protocols using biflex 3 spectrometer
1
Peptide Synthesis and Viral Quantification
2
Multimodal Characterization of Novel Materials
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High-performance liquid chromatography (HPLC) was conducted on an LC-9130 NEXT machine (Japan Analytical Industry Co., Ltd.) with toluene as the mobile phase. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was performed on a BIFLEX III spectrometer (Bruker Daltonics Inc., Germany). Vis-NIR spectra were obtained on a PE Lambda 750S spectrophotometer in CS2. Cyclic voltammograms (CV) were measured in 1,2-dichlorobenzene with 0.05 M n-Bu4NPF6 as the supporting electrolyte at a Pt working electrode with a CHI660E workstation.
3
MALDI-TOF Analysis of crChit62J4
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The crude enzyme crChit62J4 in 0.05 M Tris/HCl, pH 8, at a concentration of 1 mg/mL was subjected to matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectroscopy analysis (Supplementary Figure S3 ) with a BIFLEX III spectrometer (Bruker).
4
Spectroscopic Characterization of Organic Compounds
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All organic solvents were commercially available, dried and distilled by appropriate methods before use.
Electrospray ionization mass spectra (ESI-MS) were recorded on Waters Micromass Q-TOF mass spectrometer, MALDI-TOF MS measurements were carried out on a Bruker BIFLEX III spectrometer. Samples for C, H and N analyses were dried under vacuum and the analysis was performed with a Carlo Erba-1106 elemental analyser. 1H, 13C NMR spectra were recorded on Bruker dmx AVANCE 400 MHz, 600 MHz NMR spectrometers at room temperature with CDCl3/TDF/DMSO/CD3CN as solvent, chemical shifts are referenced to residual solvent peaks with respect to TMS = δ 0 ppm. Experimental DEPT spectra were recorded on Bruker dmx AVANCE 600 MHz NMR spectrometers for proton with flip angle of the final I-spin pulse (β) set to 3π/4.
UV-vis absorption spectra were recorded on a Hitachi U-3010 spectrophotometer. Fluorescence spectra were recorded on a Hitachi F-4500 fluorescence spectrophotometer. Fluorescence quantum yields (Фf) were determined by the absolute method on HAMAMATSU Absolute PL Quantum Yield Spectrometer C11347.
Electrospray ionization mass spectra (ESI-MS) were recorded on Waters Micromass Q-TOF mass spectrometer, MALDI-TOF MS measurements were carried out on a Bruker BIFLEX III spectrometer. Samples for C, H and N analyses were dried under vacuum and the analysis was performed with a Carlo Erba-1106 elemental analyser. 1H, 13C NMR spectra were recorded on Bruker dmx AVANCE 400 MHz, 600 MHz NMR spectrometers at room temperature with CDCl3/TDF/DMSO/CD3CN as solvent, chemical shifts are referenced to residual solvent peaks with respect to TMS = δ 0 ppm. Experimental DEPT spectra were recorded on Bruker dmx AVANCE 600 MHz NMR spectrometers for proton with flip angle of the final I-spin pulse (β) set to 3π/4.
UV-vis absorption spectra were recorded on a Hitachi U-3010 spectrophotometer. Fluorescence spectra were recorded on a Hitachi F-4500 fluorescence spectrophotometer. Fluorescence quantum yields (Фf) were determined by the absolute method on HAMAMATSU Absolute PL Quantum Yield Spectrometer C11347.
5
Synthesis and Characterization of D-Ribose Compounds
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Commercial D-ribose (Merck, New Jersey, NJ, USA) was used. All reactions were monitored by thin-layer chromatography (TLC) on Kieselgel 60 F254 Silica Gel plates (E. Merck, 0.20 mm thickness) using eluent system (v/v) 3:1 CHCl3-MeOH (Merck). The spots were detected by spraying with 5% ethanolic H2SO4 and charring. Other reagents, such as C2H5OH, H2SO4, 2-butanon, tertiary amines, CH3CN, CH2Cl2, Tf2O were obtained from Merck. 1H-NMR and 13C-NMR spectra were recorded at 25 °C with a Varian Mercury (Agilent, Santa Clara, CA, USA) spectrometer at 400.49 and 100.70 MHz, respectively, with Me4Si as the internal standard; positive-ion mode MALDITOF mass spectra was done on a Bruker Biflex III spectrometer (Billerica, Massachusetts, MA, USA).
6
Characterization of Luminescent Molecular Probes
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All reagents were obtained from
Beijing Innochem Co. and used without further purification. 1H NMR and 13C NMR spectra were recorded on Bruker AVANCE
400 spectrometer. Ultrapure water (18 MΩ/cm) was used for the
preparation of all aqueous solutions. Matrix-assisted laser desorption/ionization–time
of flight–mass spectrum was measured by a Bruker BIFLEX III
spectrometer. UV–vis absorption spectra were recorded with
a Hitachi U-3010 spectrophotometer. Photoluminescence spectra were
carried out on a Hitachi F-7000 spectrophotometer. Fluorescence quantum
yields were determined by using Nile Red (0.38 in methanol) as standards.
Beijing Innochem Co. and used without further purification. 1H NMR and 13C NMR spectra were recorded on Bruker AVANCE
400 spectrometer. Ultrapure water (18 MΩ/cm) was used for the
preparation of all aqueous solutions. Matrix-assisted laser desorption/ionization–time
of flight–mass spectrum was measured by a Bruker BIFLEX III
spectrometer. UV–vis absorption spectra were recorded with
a Hitachi U-3010 spectrophotometer. Photoluminescence spectra were
carried out on a Hitachi F-7000 spectrophotometer. Fluorescence quantum
yields were determined by using Nile Red (0.38 in methanol) as standards.
7
Thorough Characterization of Chemical Compounds
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Starting materials were purchased and used without further purification. Thin-layer chromatography (TLC) was performed on aluminium plates coated with MerckSilica gel 60 F254. When necessary, silica gel 60 (35-70 mesh, SDS) was used to isolate the desired derivatives. C NMR spectra were recorded using deuterated solvents as internal references on a BRUKER Advance DRX 300 or a BRUKER AV400 spectrometer. Multiplicities are denoted as follows: s = singlet, d = doublet, t = triplet, m = multiplet, br = broad. The mass spectra were recorded on a Q-ToF spectrometer (accurate mass measurements were achieved using sodium formate as internal reference), a Jeol JMS 700 (high-resolution mass spectra (HRMS) or a Bruker Biflex III spectrometer (MALDI-TOF). The infrared absorption spectra were recorded on an FTIR BRUKER VERTEX 70. Optical microscopy was performed by employing a Leica DM 2500P microscope. SEM micrographs were recorded by scanning electron microscopy (SEM) with a JEOL JSM 6301F operating at tensions of 3 kV.
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