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Cary 100 bio spectrophotometer

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The Cary 100 Bio spectrophotometer is a laboratory instrument designed for measuring the absorbance of light by samples. It is capable of analyzing a wide range of samples, including biological and chemical substances, across a broad spectral range.

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54 protocols using cary 100 bio spectrophotometer

1

DPPH Radical Scavenging Assay for Mdivi-1

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To measure the antioxidant activity of mdivi-1, we also performed an assay based on the inhibition of the absorbance of the radical DPPH• (Figure 1B), as previously described [31 (link)]. Mdivi-1 and Trolox were diluted from 50 mM stocks, prepared in DMSO and water, respectively, to 2x stocks in absolute ethanol. Then, 1 mL of 100 mM DPPH made in absolute ethanol was added to 1 mL of 2x mdivi-1 or Trolox to obtain the final reaction mixtures with the indicated concentrations of compounds. The reaction mixtures were vortexed vigorously for 10 s and allowed to stand at room temperature, protected from light, for 30 min. Absorbance was determined at 517 nm on a Varian Cary 100 Bio spectrophotometer. Mdivi-1 antioxidant activity was calculated as the percentage inhibition of DPPH• solution absorbance equated against a Trolox standard curve.
Bordt E.A., Zhang N., Waddell J, & Polster B.M. (2022). The Non-Specific Drp1 Inhibitor Mdivi-1 Has Modest Biochemical Antioxidant Activity. Antioxidants, 11(3), 450.
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2

Determination of Total Phenolic Index

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The total phenolic index (TPI) of the WSs was determined according to Cetó et al. [65 (link)]. Briefly, WSs were diluted with distilled water and the absorbance was measured directly at 280 nm using a Varian Cary 100 Bio spectrophotometer (Santa Clara, CA, USA) with a 10 mm quartz cuvette. TPI value of each sample was calculated by multiplying the measured absorbance by the dilution factor. The analyses were performed in triplicate.
Oliveira-Alves S., Lourenço S., Anjos O., Fernandes T.A., Caldeira I., Catarino S, & Canas S. (2021). Influence of the Storage in Bottle on the Antioxidant Activities and Related Chemical Characteristics of Wine Spirits Aged with Chestnut Staves and Micro-Oxygenation. Molecules, 27(1), 106.
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3

Cytochrome c Reduction Assay

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The parameter was assayed in homogenates of hippocampi 6 and 24 h after ACA. Samples (500 µl) of the respective tissue homogenates were fivefold frozen and defrosted to permeabilize membranes. A volume of 15 µl (40 µg protein) was used to run the standard assay at 30°C. The reduction of cytochrome c was followed by measuring the absorption at 550 nm with a Cary 100 Bio spectrophotometer (Varian). The antimycin A-sensitive absorption was used for quantification.
Keilhoff G., Esser T., Titze M., Ebmeyer U, & Schild L. (2017). Gynostemma pentaphyllum is neuroprotective in a rat model of cardiopulmonary resuscitation. Experimental and Therapeutic Medicine, 14(6), 6034-6046.
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4

Elemental Analysis by ICP-MS

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Unless otherwise noted, starting materials were obtained from commercial
suppliers and used without further purification. Water was distilled
and further purified by a Millipore cartridge system (resistivity
18 MΩ). Elemental analyses were performed by inductively coupled
plasma mass spectrometry on a Thermo Scientific XSERIES 2 ICP-MS fitted
with an ESI PC3 Peltier cooled spray chamber, SC-FAST injection loop,
and SC-4 autosampler at the Department of Geology at the University
of Minnesota. Samples were diluted appropriately and analyzed in the
presence of a 20 ppb of In internal standard using the He/H2 collision-reaction
mode. UV–vis spectra were measured with a Varian Cary 100 Bio
spectrophotometer. Data were collected between 220 and 800 nm using
a quartz cell with a path length of 10 mm. Luminescence data were
recorded on a Varian Eclipse Fluorescence spectrophotometer using
a quartz cell with a path length of 10 mm.
Peterson K.L., Dang J.V., Weitz E.A., Lewandowski C, & Pierre V.C. (2014). Effect of Lanthanide Complex Structure on Cell Viability and Association. Inorganic Chemistry, 53(12), 6013-6021.
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5

Measurement of Antioxidant Activity via ABTS Assay

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To measure the antioxidant activity of mdivi-1, we performed an assay as described in [30 (link)] that is based on the inhibition of the absorbance of the radical cation of ABTS (ABTS•+). ABTS (7 mM) was prepared in water, and ABTS•+ (Figure 1A) was produced by combining ABTS with 2.45 mM potassium persulfate and allowing the mixture to react at room temperature in the dark for 16–24 h prior to experimental use. Trolox standard and mdivi-1 compound were mixed with ABTS•+, and absorbance at 734 nm was determined on a Varian Cary 100 Bio spectrophotometer (Walnut Creek, CA, USA) after a 6 min incubation. The antioxidant activity of mdivi-1 was calculated as the percentage inhibition of ABTS•+ solution absorbance equated against a Trolox standard curve.
Bordt E.A., Zhang N., Waddell J, & Polster B.M. (2022). The Non-Specific Drp1 Inhibitor Mdivi-1 Has Modest Biochemical Antioxidant Activity. Antioxidants, 11(3), 450.
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6

Multimodal Analytical Characterization

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1H NMR spectra were recorded on a Varian 300 MHz spectrometer at room temperature and referenced to a residual deuterated solvent. UV-vis spectra were recorded in a 1 cm quartz cuvette on a Varian Cary 100 Bio spectrophotometer. ATR-FTIR spectra were recorded with an Agilent/Cary 630 FTIR KBr or ZnSe engine. The electrochemical data were obtained using CH Instrument 600E Electrochemical workstation. ESI-Mass spectral data were collected using Agilent Technologies 6530 Accurate-Mass Q-TOF LC/MS equipped with a Jet Stream electrospray ionization (ESI) source. Elemental analyses were performed by Galbraith Atlantic Microlabs, Norcross, GA.
Blade G.A., Parveen R., Jaimes J.L., Ilustre W., Saldaña D., Ivan D.A., Lynch V.M., Cundari T.R, & Toledo S. (2020). A family of structural and functional models for the active site of a unique dioxygenase: Acireductone Dioxygenase (ARD). Journal of inorganic biochemistry, 212, 111253.
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7

Lysozyme Aggregation Monitored by Spectroscopy

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Denaturing and non-denaturing electrophoresis were performed as described previously [39 (link)]. The aggregation of fluorescein-labeled lysozyme was initiated by the addition of 10 mM DTT to 10 µM protein solution in 20 mM Tris-HCl, pH 7.5, 10 mM MgCl2 both in the absence and in the presence of equimolar GroEL, GroES, and 10 mM ADP. After certain incubation time, the mixtures were centrifugated for 10 min at 12,000× g and fluorescence spectra of supernatants were measured. The protein solution without DTT was used as a standard for fluorescence intensity. Fluorescence spectra and anisotropy, as well as bioluminescence, were measured using a Cary Eclipse spectrofluorimeter (Varian Medical Systems, Palo Alto, CA, USA). Static and manual-mixing kinetic measurements were performed in a standard 1 × 1 × 4 cm quartz cell using a magnetic stirrer. A stopped-flow device combined with a Chirascan spectropolarimeter (Applied Photophysics Ltd, London, UK) was used for the registration of fast kinetics. The approximation of the kinetic curves was made using the SigmaPlot computer program (Systat Software Inc., Chicago, IL, USA).
The absorption spectra were recorded using a Cary 100 Bio spectrophotometer (Varian Medical Systems, Palo Alto, CA, USA).
Marchenkov V., Gorokhovatsky A., Marchenko N., Ivashina T, & Semisotnov G. (2020). Back to GroEL-Assisted Protein Folding: GroES Binding-Induced Displacement of Denatured Proteins from GroEL to Bulk Solution. Biomolecules, 10(1), 162.
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8

Characterization of Cross-Linked Gold Nanoparticles

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Dynamic light scattering (DLS, 90 plus Brookhaven Instruments Corporation, USA) was used for size determination. The single and intercross-linked GNPs images were obtained by PHILIPS CM30 Scanning Transmission Electron Microscope (Philips Electron Optics, Eindhoven, The Netherlands). For UV-Visible spectroscopy, the Cary 100 bio spectrophotometer (Varian, Australia) was used. Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR- FTIR) was performed using Nicolet Nexus 470 FTIR (Nicolet, Madison, WI, USA) spectrometer. IrAnalyze software was also applied for IR spectrum interpretation (LabCognition, Ft. Myers, FL). TMicro centrifuge (Sigma, USA) was used to separate GNPs at different conditions. The samples were centrifuged at 14000 rpm for 30 min at 4 °C. The chemiluminescence emissions were recorded at 425 nm by fluorescence micro plate reader (H4, Bio Tech Co, USA).
Torabi R, & Ghourchian H. (2020). Ultrasensitive nano-aptasensor for monitoring retinol binding protein 4 as a biomarker for diabetes prognosis at early stages. Scientific Reports, 10, 594.
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9

UV Melting Analysis of DNA Samples

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UV melting experiment was carried out on a Varian CARY-100 BIO Spectrophotometer. The low concentration experiment was carried out at 295 nm using a 1 cm cell. The concentration of DNA was adjusted to the absorbance at 295 nm in the range between 0.4 and 0.8. In the high concentration experiment, NMR samples were used directly in a 1 mm cell and the wavelengths were adjusted between 300 and 305 nm to obtain an absorbance of less than 1.0. The temperature was increased/decreased from 10 to 80°C with the rate of 0.5°C/min. Melting temperatures were determined from the first order derivatives of melting profiles.
Podbevšek P, & Plavec J. (2015). KRAS promoter oligonucleotide with decoy activity dimerizes into a unique topology consisting of two G-quadruplex units. Nucleic Acids Research, 44(2), 917-925.
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10

Singlet Oxygen Generation by ICG-AuNP Clusters

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ICG-AuNP clusters and various controls were examined for their ability to generate reactive oxygen species, using the reporter reagent Singlet Oxygen Sensor Green. SOSG-containing solutions (2.5 mL, 10 μM SOSG) were added to a quartz cuvette and mixed continuously by magnetic stirrer for 30 minutes. Aliquots (60 μL) were withdrawn at each of the following timepoints: 0, 1, 5, 10, 20, and 30 minutes. Samples were then diluted 20-fold in water, and fluorescence was read by fluorimeter (ex = 488 nm; em = 523 nm) and normalized to the 0-minute reading. Solutions of ICG-AuNP clusters and free ICG (0.015 mg mL−1) were tested with or without sodium azide (10 mM). Samples were either kept in the dark or were irradiated continuously for 30 minutes (808 nm; 1.2 W power, 0.1 cm2 area). Irradiated samples of clusters and ICG also had surface temperature monitored using a FLIR ONE thermal imaging camera (FLIR Systems, Wilsonville, OR); based on this data, a subset of samples were heated with equivalent temperature and timing under dark conditions (heat generated by Varian-Cary 100 Bio spectrophotometer, thermal accessory). Sample conditions generating greater than 1.5-fold change in SOSG fluorescence were performed in triplicate; all others were performed as single assays.
Higbee-Dempsey E., Amirshaghaghi A., Case M.J., Miller J., Busch T.M, & Tsourkas A. (2019). Indocyanine Green–Coated Gold Nanoclusters for Photoacoustic Imaging and Photothermal Therapy. Advanced therapeutics, 2(9), 1900088.
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