8453 spectrometer

Manufactured by Agilent Technologies

The 8453 spectrometer is a compact and versatile UV-Vis spectrophotometer designed for a wide range of analytical applications. It features a diode array detector that provides rapid and precise measurements across the ultraviolet and visible light spectrum. The 8453 spectrometer is capable of performing various spectroscopic analyses, including quantitative determinations, reaction monitoring, and basic research.

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Lab products found in correlation

300 mhz spectrometer, by Bruker (2 mentions) Rf 5301pc spectrophotofluorimeter, by Shimadzu (2 mentions) 713 ph meter, by Metrohm (2 mentions) J 810 spectropolarimeter, by Jasco (1 mentions) Gcms qp2020 gas chromatograph mass spectrometer, by Shimadzu (1 mentions) Avance drx 500 spectrometer, by Bruker (1 mentions) F 4500 fluorimeter, by Hitachi (1 mentions) Fls980, by Edinburgh Instruments (1 mentions) Phi 5000 versaprobe spectrometer, by Physical Electronics (1 mentions) Jem arm200f, by JEOL (1 mentions) Pd 10 column, by GE Healthcare (1 mentions) Synergy plate reader, by Agilent Technologies (1 mentions) Autoflex mass spectrometer, by Bruker (1 mentions) Dionex ics 5000 instrument, by Thermo Fisher Scientific (1 mentions) Sx 20 instrument, by Applied Photophysics (1 mentions) Emxplus spectrometer, by Bruker (1 mentions) Av 400 spectrometer, by Bruker (1 mentions) Fp 6500 spectrophotometer, by Jasco (1 mentions) Aviii 400 mhz nmr spectrometer, by Bruker (1 mentions) Vertex 70 ftir spectrometer, by Bruker (1 mentions)

29 protocols using 8453 spectrometer

Gdn·HCl-induced Unfolding of A15C Ngb

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Gdn·HCl-induced unfolding of A15C Ngb was studied by Agilent 8453 spectrometer. 10 μL of 2 mM protein sample was added into 2 mL of a guanidine hydrochloride (Gdn·HCl) solution (pH 7.0) with various concentrations (0–6.0 M). After incubation for 30 min, the UV-Vis spectrum was recorded. Control experiment was performed for WT Ngb under the same conditions. The unfolding transition was monitored at different Gdn·HCl concentrations through the changes of the absorbance of the Soret band. The denaturation midpoint (C_m) was calculated by fitting the absorbance of Soret band versus the concentration of Gdn·HCl to a two-state Boltzmann function (eqn (1)). Here, A is the absorbance of Soret band; A₁ and A₂ are the initial and final absorbance of Soret band, respectively; C is the concentration of Gdn·HCl; C_m is the concentration of Gdn·HCl at midpoint of denaturation.

Liu H.X., Li L., Yang X.Z., Wei C.W., Cheng H.M., Gao S.Q., Wen G.B, & Lin Y.W. (2019). Enhancement of protein stability by an additional disulfide bond designed in human neuroglobin. RSC Advances, 9(8), 4172-4179.

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Spectroscopic Analysis of Biomolecules

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UV–Vis spectra were collected using an Agilent 8453 spectrometer, with samples contained in 1 cm × 1 cm quartz cuvettes. Each spectrum was taken with reference to a blank solvent spectrum. Circular dichroism spectra were collected on the same samples using a Jasco J-810 spectropolarimeter with a scan speed of 100 nm/min and bandwidth of 1 nm; the spectra were generally strong enough that signal averaging was unnecessary. As in the UV–vis measurements, a blank toluene baseline spectrum was subtracted from the CD spectra before analysis.

Dunlap-Shohl W.A., Tabassum N., Zhang P., Shiby E., Beratan D.N, & Waldeck D.H. (2024). Electron-donating functional groups strengthen ligand-induced chiral imprinting on CsPbBr3 quantum dots. Scientific Reports, 14, 336.

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Comprehensive Analytical Characterization of Toxicant Metabolites

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¹H NMR spectra were obtained using a Bruker 300 MHz spectrometer. UV-Visible spectra were obtained using an Agilent 8453 spectrometer equipped with a photodiode array detector. Fluorescence spectra were obtained using a Shimadzu RF-5301PC spectrophotofluorimeter. All GCMS measurements were obtained using a Shimadzu GCMS-QP2020 gas chromatograph-mass spectrometer. All toxicants and toxicant metabolites (compounds 1–10, Figure 1) were purchased from Sigma Aldrich and used as received. Fluorophore 11 was synthesized following literature-reported procedures.^{46 (link)} Fluorophores 12 and 13 were purchased from Sigma Aldrich and used as received.

DiScenza D.J., Lynch J., Verderame M., Serio N., Prignano L., Gareau L, & Levine M. (2017). Efficient Fluorescence Detection of Aromatic Toxicants and Toxicant Metabolites in Human Breast Milk. Supramolecular chemistry, 30(4), 267-277.

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Synthesis and Characterization of Neurotransmitter Probes

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The indicators i1, i2 and i3 were synthesised as described in our earlier papers.^{33,34 (link)} The synthesis of WP6 (ammonium salt) was carried out as reported^{56–58 (link)} with slight modifications in the procedure and described in the ESI† in detail. The starting materials and solvents were purchased from commercial suppliers and used without further purification. The neurotransmitters were obtained from Sigma (choline and acetylcholine as chloride salts, histamine as dihydrochloride, serotonin and dopamine as hydrochloride).
The absorption and fluorescence spectroscopic experiments were carried out in HEPES buffers of pH 7.4. The fluorescence quantum yields were determined using Coumarin 153 as reference. The absorption spectra were recorded on an Agilent 8453 spectrometer. The fluorescence spectra were measured on a PerkinElmer 50B spectrofluorimeter. The NMR spectra were acquired on a 500 MHz Bruker Avance DRX-500 spectrometer. All the spectral measurements were carried out at 25 °C.

Paudics A., Kubinyi M., Bitter I, & Bojtár M. (2019). Carboxylato-pillar[6]arene-based fluorescent indicator displacement assays for the recognition of monoamine neurotransmitters. RSC Advances, 9(29), 16856-16862.

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AuNPs Aggregation in Sodium Halides

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The as-synthesized 13 nm AuNPs (particle concentration ∼13 nM) were first diluted with an equal volume of 10 mM of the various buffers and then equilibrated at 23 °C for 1 h before titration. For each sample, 20 μL of sodium halide was quickly mixed with 80 μL of 6.5 nM AuNPs and incubated for 1 min. The UV-vis extinction spectra were then measured using an Agilent 8453 spectrometer. The extinction values at 520 nm and 650 nm were recorded.

Jimmy Huang P.J., Yang J., Chong K., Ma Q., Li M., Zhang F., Moon W.J., Zhang G, & Liu J. (2020). Good's buffers have various affinities to gold nanoparticles regulating fluorescent and colorimetric DNA sensing. Chemical Science, 11(26), 6795-6804.

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Enzymatic Nitric Oxide Quantification

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UV-vis absorbance spectra were recorded on Agilent 8453 spectrometer for eNOS_oxy concentration determination. The activity of eNOS_oxy was determined using a Spectra max plus 384 plate reader based on the Griess reaction through a standard calibration curve. It quantifies NO in the form of nitrite (NO₂^-) in a two-step diazotization reaction in which acidified nitrite produces a nitrosating agent, which is then derivatized to produce the final azo-product with a maximum absorption at 540 nm.
Catalysis of nitric oxide production (in the form of nitrite) from N-hydroxyarginine (NOHA) and H₂O₂ by eNOS_oxy and eNOS_oxy bound nanodisc were assayed in 96-well microplates at 37 °C as described previously with modifications. Assays (50 μl final volume) contained 20 mM Tris, 100 mM NaCl, pH 7.4, 500 nM eNOS_oxy, 1 mM NOHA, 0.5 mM DTT, 30 mM H₂O₂, and 10 μM H₄B. Reactions were started by adding H₂O₂ and stopped after 10 min by adding catalase (1300 units). Griess reagents 50 μl of sulfanilamide was added and incubated for 10 min in dark followed by 50 μl of NED which was treated the same. Finally, the assay plate was read at 540 nm in a Spectra max plate reader. Nitrite production was quantitated based on NaNO₂ standards.

Altawallbeh G., Haque M.M., Streletzky K.A., Stuehr D.J, & Bayachou M. (2017). Endothelial Nitric Oxide Synthase Oxygenase on Lipid Nanodiscs: A Nano-assembly Reflecting Native-Like Function of eNOS. Biochemical and biophysical research communications, 493(4), 1438-1442.

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UV–Vis Spectroscopy Protocol

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UV–vis
absorption
spectra were recorded with an Agilent 8453 spectrometer. Solvents
were of spectroscopic or equivalent grade. The pH of samples was measured
with a Metrohm 713 pH meter, and adjustments of the hydrogen ion concentration
were made with diluted HCl and NaOH solutions.

Martínez-Camarena Á., Savastano M., Blasco S., Delgado-Pinar E., Giorgi C., Bianchi A., García-España E, & Bazzicalupi C. (2021). Assembly of Polyiodide Networks with Cu(II) Complexes of Pyridinol-Based Tetraaza Macrocycles. Inorganic Chemistry, 61(1), 368-383.

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Spectroscopic Characterization of Organometallic Complexes

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IR spectra were recorded on a Mattson RS FTIR instrument, averaging 64 scans at resolution 2 cm^–1. ATR-IR spectra were an average of 32 scans. UV/visible absorption spectra were measured using an Agilent 8453 spectrometer. Steady state emission spectra were measured using a Hitachi F-4500 fluorimeter. The fluorescence was taken against a ZnTPP reference for the bromide complexes and against the individual dyad ligand ZnTPP-link-Bpy for the picoline complexes. Time-resolved emission was measured with an Edinburgh Instruments FLS980 equipped with a 560 nm pulsed LED (EPLED 560, pulsewidth 1.5 ns) and a red PMT detector. All samples were either degassed by three freeze–pump–thaw cycles or de-aerated by purging the sample with Ar. Correction was applied for instrument response. All absorption and emission measurements were made in 10 × 10 mm quartz cuvettes.

Windle C.D., George M.W., Perutz R.N., Summers P.A., Sun X.Z, & Whitwood A.C. (2015). Comparison of rhenium–porphyrin dyads for CO2 photoreduction: photocatalytic studies and charge separation dynamics studied by time-resolved IR spectroscopy †Electronic supplementary information (ESI) available: NMR spectra, cyclic voltammograms, crystallographic data, fluorescence data, spectra from photoreactions and photocatalytic data. CCDC 1406000 and 1406001. For ESI and crystallographic data in CIF or other electronic format see DOI: 10.1039/c5sc02099a. Chemical Science, 6(12), 6847-6864.

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Synthesis and Characterization of Compound 10

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All chemicals were obtained from Sigma Aldrich Chemical Company and were used as received. Compound 10 was synthesized following literature-reported procedures (21 (link)). Human plasma was obtained from Innovative Technologies. Coconut water was obtained from CVS Pharmacy in Kingston, Rhode Island. ¹H NMR spectra were recorded on a Bruker 300 MHz spectrometer. UV-Visible spectra were recorded on an Agilent 8453 spectrometer equipped with a photodiode array detector. Fluorescence spectra were recorded on a Shimadzu RF-5301PC spectrophotofluorimeter.

Serio N., Prignano L., Peters S, & Levine M. (2014). Detection of Medium-Sized Polycyclic Aromatic Hydrocarbons via Fluorescence Energy Transfer. Polycyclic aromatic compounds, 34(5), 561-572.

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Characterization of Nanomaterials by Advanced Spectroscopy

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UV-vis absorption spectra were recorded on an Agilent 8453 spectrometer. ESI-MS was conducted on a Bruker Impact II time-of-flight MS system. TEM images and HAADF-STEM images were obtained with a JEM-2100UHR field emission electron microscope at an accelerating voltage of 200 kV. Atomic resolution high magnification STEM image was obtained using an aberration-corrected transmission electron microscope (JEOL, JEM-ARM200F) operated at 200 kV. XRD patterns were collected on a D8 Advance X-ray powder diffractometer equipped with a Cu Kα radiation source (λ = 0.15405 nm) at 40 kV. All the diffraction data were collected in a 2θ range from 5° to 75° at a scanning rate of 8° min⁻¹. XPS analyses were acquired with a PHI 5000 Versa Probe spectrometer from ULVACPHI using an Al Kα (1486.6 eV) photon source. All binding energies were calibrated using the C(1s) carbon peak (284.8 eV), which was applied as an internal standard. TGA was performed on SDT650 from NETZSCH. The starting temperature was 20 °C with a 5 °C min⁻¹ ramp rate to 800 °C under 100 mL min⁻¹ airflow⁵⁵.

Wang X., Zhao L., Li X., Liu Y., Wang Y., Yao Q., Xie J., Xue Q., Yan Z., Yuan X, & Xing W. (2022). Atomic-precision Pt6 nanoclusters for enhanced hydrogen electro-oxidation. Nature Communications, 13, 1596.

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