Optical rotations were measured with a Jasco p-1020 digital polarimeter. UV spectra were recorded on a Shimadzu 2401PC spectrophotometer. IR spectra were obtained on a Bruker Tensor 27 infrared spectrophotometer with KBr pellets. 1H, 13C and 2D NMR spectra were obtained on Bruker AV-600, AVANCE III-500 and 400 MHz spectrometers with SiMe4 as an internal standard. Chemical shifts (δ) were expressed in ppm with reference to the solvent signals. MS data were recorded on an UPLC-IT-TOF MS. Column chromatography (CC) was performed on either silica gel (200–300 mesh, Qingdao Marine Chemical Co., Ltd., Qingdao, China) or RP-18 silica gel (20–45 μm, Fuji Silysia Chemical Ltd., Japan). Fractions were monitored by TLC on silica gel plates (GF254, Qingdao Marine Chemical Co., Ltd., Qingdao, China), and spots were visualized with Dragendorff’s reagent spray. MPLC was performed using a Buchi pump system coupled with RP-18 silica gel-packed glass columns (15 × 230 and 26 × 460 mm, respectively). HPLC was performed using Waters 1525 pumps coupled with analytical or preparative Sunfire C18 columns (4.6 × 150 and 19 × 250 mm, respectively). The HPLC system employed a Waters 2998 photodiode array detector and a Waters fraction collector III.
2401pc spectrophotometer
Manufactured by Shimadzu
Sourced in Japan
The 2401PC spectrophotometer is a compact and versatile instrument designed for a wide range of spectroscopic applications. It features a high-resolution monochromator and a photomultiplier tube detector, providing accurate and reliable absorbance measurements across the visible and near-ultraviolet wavelength range. The instrument is capable of performing tasks such as kinetic studies, concentration determination, and spectrum scanning.
Automatically generated - may contain errors
Lab products found in correlation
P 1020, by Jasco (2 mentions)
Silica gel, by Qingdao Marine Chemical (2 mentions)
Uv 240, by Shimadzu (2 mentions)
1525 pump, by Waters Corporation (1 mentions)
2998 photodiode array detector, by Waters Corporation (1 mentions)
Tensor 27 infrared spectrophotometer, by Bruker (1 mentions)
Fraction collector 3, by Waters Corporation (1 mentions)
Avance 3 500, by Bruker (1 mentions)
Agilent g6230 tof mass spectrometer, by Agilent Technologies (1 mentions)
Ultimate 3000, by YMC (1 mentions)
P 1020 polarimeter, by Jasco (1 mentions)
Avance 3 500 mhz spectrometer, by Bruker (1 mentions)
Hplc grade methanol, by Tedia (1 mentions)
Acetonitrile, by Tedia (1 mentions)
Pack ods a column, by YMC (1 mentions)
No 1 and 3 mm papers, by Cytiva (1 mentions)
Prism 6, by GraphPad (1 mentions)
Lc msd tof, by Agilent Technologies (1 mentions)
Unity 300 mhz, by Agilent Technologies (1 mentions)
Omni flex, by Bruker (1 mentions)
14 protocols using 2401pc spectrophotometer
1
Comprehensive Spectroscopic Characterization
2
Spectroscopic Analysis of Chemical Compounds
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Optical rotations were measured in MeOH on a P-1020 polarimeter (JASCO, Tokyo, Japan). UV spectra were obtained on a 2401PC spectrophotometer (Shimadzu Co., Tokyo, Japan), while 1D and 2D NMR spectra were recorded on a Bruker AVANCEIII-500 MHz spectrometer (Bruker, Ettlingen, Germany) in acetone-d6, CD3OD, and C5D5N using tetramethylsilane (TMS) as an internal reference standard. Chemical shifts were expressed as δ in ppm and the coupling constants (J) were given in Hz. High-resolution electrospray mass spectroscopy was performed on an Agilent G6230 TOF mass spectrometer (HR-ESI-MS) (Agilent Technologies, California, CA, USA) and a Waters Autospec Premier 776 mass spectrometer (HR-EI-MS) (Waters Technologies, Massachusetts, MA, USA). High-performance liquid chromatography (HPLC) was conducted on an Ultimate 3000 HPLC system (Dionex Co., Sunnyvale, CA, USA) equipped with an Ultimate 3000 pump and variable wavelength detector, as well as a semi-preparative YMC-Pack ODS-A column (250 × 10 mm, 5 µm, YMC, Kyoto, Japan). Column chromatography (CC) was conducted over silica gel (300–400 mesh, Qingdao Haiyang Co. Ltd., Qingdao, China). HPLC grade methanol and acetonitrile were purchased from Tedia Co. Inc, (Fairfield, OH, USA).
3
Characterization of γ-PGA by NMR, APC, FT-IR, and SEM
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NMR spectra were acquired on DELTA 300 MHz equipment for 1H NMR (Bruker, Billerica, MA, USA). For this analysis, 10 mg of γ-PGA was dissolved in D2O under an inert atmosphere. The molecular weight was determined by APC with a Waters UPLC, Agilent (Santa Clara, CA, USA) equipped with a UV detector operated at 215 nm with a silicon column and an aqueous solution of NaHPO4:acetonitrile at a ratio of 4:1 as the eluent. ATR Fourier transform infrared (FT-IR) analysis was carried out with a Nicolet iS5 Thermo Scientific iD7ATR (Waltham, MA, USA) ranging from 4000–600 cm−1 at a resolution of 16 cm−1 with 64 scans, using germanium as the reference standard material. SEM characterization was performed with a JEOL JSM-7041F (Akishima, Tokyo, Japan). The samples were gold-palladium sputter-coated, the size distribution and porous size was determined with ImageJ software version 1.43 (Maryland, USA). Absorption spectra were obtained with a Shimadzu 2401PC spectrophotometer (Shimadzu corporation, Kyoto, Japan).
4
Spectroscopic Characterization of Organic Compounds
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UV spectra were recorded on Shimadzu model UV-240 and 2401 PC spectrophotometer (Shimadzu Inc., Tokyo, Japan). NMR experiments were recorded on Bruker spectrometer (Switzerland) 600 (1H NMR spectra: 600 MHz; 13C NMR spectra: 150 MHz). The chemical shifts are given in δ (ppm) relative to tetramethylsilane (Me4Si) TMS. Column chromatography (CC) was carried out on silica gel, 60–200 mesh, Fluca, India) and Sephadex LH-20 (Pharmazia, Uppsala, Sweden). Paper chromatography (PC, descending) Whatman No. 1 and 3 mm papers, silica gel aluminum sheets (20 × 20) using solvent systems hexane: chloroform: methanol 90:8:2 (S-1), 15% HOAc(S-2), BAW (n-BuOH: HOAc: H2O) 3:1:1(S-3), and BAW 4:1:5, the upper layer (S-4).
5
LSD1 Activity Assay Protocol
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LSD1 activity assays were performed using a peroxidase-coupled assay under aerobic conditions as described previously (Forneris et al. 2005 (link)). A 150 µL reaction mixture contained 50 mM HEPES(Na), pH 7.5, 0.2–0.3 µM LSD1, 1 µg of HRP, 0.1 mM 4-aminoantipyrine, 1.0 mM 3,5-dichloro-2-hydroxybenzenesulfonic acid and substrate concentrations as indicated in the main text. Enzyme reactions were initiated by the addition of substrate (dimethyl-Lys4 Histone H3 peptide aa 1–21 [diMeK4H31–21] [Sigma]) and the time course of the reaction was monitored using a Shimadzu 2401-PC spectrophotometer (λ = 515 nm) at 25°C in a thermostated quartz cell chamber. Initial velocity values were measured using an extinction coefficient of 26,000 M−1 cm−1 (Forneris et al. 2005 (link)). Kinetic data were fitted to a Michaelis–Menten equation using GraphPad Prism 6.0 (GraphPad Software, Inc., http://www.graphpad.com ).
6
Synthesis and Photophysical Characterization of Fluorescent Compounds
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All starting materials were purchased from Sigma-Aldrich Chemical Company. Solvents were used without further purification. Melting points were determined on an Electrothermal Mel-Temp apparatus and are not corrected. Nuclear magnetic resonance experiments as 1H and 13C NMR spectra were recorded on a Bruker advance DPX 400. Chemical shifts (ppm) are relative to (CH3)4Si for 1H and 13C. High-resolution mass spectra (HRMS) were acquired by LC/MSD TOF on an Agilent Technologies instrument with APCI as the ionization source. UV-vis absorption spectra were measured on a Shimadzu 2401 PC spectrophotometer. The emission spectra have been recorded with a Fluorolog 3 spectrofluorometer, by exciting 10 nm below the longer wavelength absorption band. Fluorescence quantum yields in solution (ϕ) were determined according to the procedure reported in the literature29 (link) and using quinine sulphate in H2SO4 0.1 M (ϕ = 0.54 at 310 nm) as the standard. Measurements were carried out by controlling the temperature at 25.0 ± 0.5 °C with a water circulating bath. Five solutions with absorbance at an excitation wavelength lower than 0.1 were analyzed for each sample and the quantum yield was averaged.
7
Spectroscopic Characterization of Compounds
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1H and 13C NMR spectra were recorded on a Varian Unity-300 MHz with tetramethylsilane (TMS) as an internal reference. Infrared (IR) spectra were measured on a Nicolet FT-SSX spectrophotometer (Bruker, Billerica, MA, USA). Elemental analysis was determined by Galbraith Laboratories, INC Knoxville (Sycamore, MO, USA). FAB+ mass spectra were taken on a JEOL JMS AX505 HA instrument (JEOL manufacturers, Tokyo, Japan). Electrospray mass spectra were taken on a Bruker Daltonic, Esquire 6000 (Bruker, Billerica, MA, USA). Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectra were taken on a Bruker Omni FLEX using 9-nitroanthracene (9NA) as a matrix. The ultraviolet–visible (UV-Vis) absorption spectra were obtained at room temperature with a Shimadzu 2401 PC spectrophotometer (Shimadzu, Kyoto, Japan).
8
Comprehensive Analytical Characterization
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Optical rotations were obtained with a JASCO P-1020 digital polarimeter, using MeOH as solvent. UV spectra were taken on Shimadzu 2401PC spectrophotometer. CD spectra were obtained by a Chirascan instrument. 1H, 13C and 2D NMR experiments were measured on Bruker DRX-500 and Avance III-600 MHz spectrometers (Bruker, Zürich, Switzerland) with the solvent signal as internal reference. Mass spectra were collected from a VG Autospec-3000 spectrometer. BRUKER Tensor-27 instrument were used to record infrared spectra (with KBr pellets). X-ray diffraction was realized on a Bruker SMART APEX CCD crystallography system. Precoated TLC plates (200–250 μm thickness, silica gel 60 F254, Qingdao Marine Chemical, Inc.) were used for the thin-layer chromatography. Semipreparative HPLC was taken on an Agilent 1100 liquid chromatography, the column used was an YMC-Pack 10 mm × 250 mm column (Pro C18 RS). Column chromatography (cc) was performed on silica gel (200–300 mesh; Qingdao Marine Chemical Inc., P. R. China), on C-18 silica gel (40–60 μm; Merck), and on Sephadex LH-20 (Amersham Pharmacia, Sweden).
9
Quantifying Polyelectrolyte Complex Hydration
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The mass of the systems in vials was weighed
at several points: empty, filled with all components, with the supernatant
removed, and dried. By comparing the mass of the systems with the
supernatant removed and when dried, we obtain the water content of
the PEC. Drying is verified via evaluation of the cobalt color; hydrated
wet cobalt is bright pink, while dehydrated cobalt is dark blue. Sufficient
cobalt was present after drying to visually verify successful dehydration.
The concentration of Co2+ ions in the supernatant phase
is determined using a Shimadzu 2401PC spectrophotometer via analysis
of the absorbance at a wavelength of 510 nm. The Co2+ concentrations
in the supernatant are compared to a polyelectrolyte-absent control
with the same initial CoCl2 concentration. The partitioning
coefficient KCo2+ is then determined
as where is the concentration of Co2+ in the supernatant, normalized for the polyelectrolyte-absent control C, which is defined as 100%. The mass of the hydrated complex
is mpec, while msup is the mass of the supernatant. An assumption was made
that the density of (hydrated) PEC was approximately equal to that
of water.40 (link)
at several points: empty, filled with all components, with the supernatant
removed, and dried. By comparing the mass of the systems with the
supernatant removed and when dried, we obtain the water content of
the PEC. Drying is verified via evaluation of the cobalt color; hydrated
wet cobalt is bright pink, while dehydrated cobalt is dark blue. Sufficient
cobalt was present after drying to visually verify successful dehydration.
The concentration of Co2+ ions in the supernatant phase
is determined using a Shimadzu 2401PC spectrophotometer via analysis
of the absorbance at a wavelength of 510 nm. The Co2+ concentrations
in the supernatant are compared to a polyelectrolyte-absent control
with the same initial CoCl2 concentration. The partitioning
coefficient KCo2+ is then determined
as where is the concentration of Co2+ in the supernatant, normalized for the polyelectrolyte-absent control C, which is defined as 100%. The mass of the hydrated complex
is mpec, while msup is the mass of the supernatant. An assumption was made
that the density of (hydrated) PEC was approximately equal to that
of water.40 (link)
10
Turbidity Measurement of PNMS Samples
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Turbidity measurement of single or mixed blank PNMS was performed by determination of %T using a UV-Visible 2401 PC spectrophotometer (Shimadzu Co., Kyoto, Japan) at 520 nm using distilled water as a blank. All PNMS were stabilized at room temperature before %T measurement (Kulthe et al., 2011 (link); Sayed et al., 2017 ).
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