All spectroscopic experiments were conducted in 50 mM, pH 7.4 Tris-HCl buffer containing 100 mM sodium chloride. CD and UV absorption spectra were recorded on a JASCO J-715 spectropolarimeter at 25 ± 0.2 °C. Temperature control was provided by a Peltier thermostat equipped with magnetic stirring. CD/UV titration experiments were performed in a rectangular quartz cell of 1 cm optical path length (Hellma, USA). Each spectrum represents the average of two scans obtained by collecting data at a scan speed of 200 nm/min. Absorption spectra were obtained by conversion of the high voltage (HT) values of the photomultiplier tube of the CD equipment into absorbance units. CD and UV curves of surfen-CDX and CDX-surfen mixtures were corrected by blank buffer or buffer solutions of CDXs, respectively. JASCO CD spectropolarimeters record CD data as ellipticity ('') in units of millidegrees (mdeg).
Cd spectropolarimeter
Manufactured by Jasco
Sourced in Japan, United States
The CD spectropolarimeter is a scientific instrument used to measure the circular dichroism (CD) of a sample. CD refers to the difference in absorption of left-handed and right-handed circularly polarized light by a sample, which can provide information about the structure and conformation of chiral molecules, such as proteins and nucleic acids.
Automatically generated - may contain errors
Lab products found in correlation
Rectangular quartz cell, by Hellma (2 mentions)
J 715 spectropolarimeter, by Jasco (2 mentions)
Lps o111 b4, by Merck Group (2 mentions)
S aureus lta, by Merck Group (2 mentions)
Copper 2 chloride, by Merck Group (1 mentions)
Folin ciocalteu, by Merck Group (1 mentions)
Tris hcl, by Merck Group (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
Phenylmethylsulfonyl fluoride pmsf, by Merck Group (1 mentions)
Calf thymus dna, by Sisco Research Laboratories (1 mentions)
Sodium hydroxide (naoh), by Sisco Research Laboratories (1 mentions)
Trichloroacetic acid, by Merck Group (1 mentions)
Glacial acetic acid, by Merck Group (1 mentions)
Nano3, by Merck Group (1 mentions)
Congo red, by Merck Group (1 mentions)
15 protocols using cd spectropolarimeter
1
Spectroscopic Characterization of Surfen-CDX
2
Spectroscopic Analysis of Drug-GAG Interactions
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Spectroscopic experiments were conducted in ultra-pure deionized water (18.2 mΩ) or in 50 mM, pH 7.0/7.4 phosphate buffer (80/140 mM NaCl) using a rectangular quartz cell of 1 cm optical path length (Hellma, USA). CD and absorption spectra were recorded on a JASCO J-715 spectropolarimeter at 25 ± 0.2 °C and represent the average of three scans obtained by collecting data at a scan speed of 100 nm/min. Temperature control was provided by a Peltier thermostat equipped with magnetic stirring. Absorption spectra were obtained by conversion of the high voltage (HT) values of the photomultiplier tube of the CD equipment into absorbance units. CD and UV curves of drug-GAG mixtures were corrected by blank water or buffer solution. JASCO CD spectropolarimeters record CD data as ellipticity ('Θ') in units of millidegrees (mdeg). The quantity of 'Θ' is converted to molar circular dichroic absorption coefficient (∆ε in M -1 cm -1 ) using the equation ∆ε = Θ/(33982cl), where, 'c' is the molar concentration of the ligand (mol/L), and 'l' is the optical pathlength expressed in cm.
3
Purification and Characterization of Biomolecules
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Tnz was purchased
from Sigma-Aldrich
and purified by re-crystallization from methanol. Copper(II) chloride
(CuCl2·2H2O), copper(II) acetate [Cu(OAc)2·H2O], NaCl, NaNO3, trichloroacetic
acid (TCA), glacial acetic acid, sodium dihydrogen phosphate, disodium
hydrogen phosphate, anthrone as reagent, Folin-Ciocalteu as reagent,
congo red, cetyltrimethylammonium bromide (CTAB), chitin flakes, Tris–HCl,
β-mercaptoethanol, phenylmethylsulfonyl fluoride (PMSF), and
KCl (all AR grade) were purchased from E. Merck, India. Thymine, cytosine,
and adenine were purchased from TCI, Japan, and calf thymus DNA, crystal
violet (CV), ethyl acetate, hydroxyl amine, NaOH, and ferric chloride
were procured from Sisco Research Laboratories, India. Calf thymus
DNA was dissolved in triple distilled water in the presence of 120
mM NaCl, 35 mM KCl, and 5 mM MgCl2. Its concentration was
determined using a molar extinction coefficient of 6600 M–1 cm–1 at 260 nm. Absorbance of the DNA solution
was also measured at 280 nm; A260/A280 was determined. The value found in the range
1.8–1.9 was considered ready for use, not requiring further
purification. Quality of calf thymus DNA was verified using circular
dichroism (CD), recording its response at 260 nm on a CD spectropolarimeter
(J815—JASCO, Japan). Aqueous solutions of all other substances
were prepared in triple distilled water.
from Sigma-Aldrich
and purified by re-crystallization from methanol. Copper(II) chloride
(CuCl2·2H2O), copper(II) acetate [Cu(OAc)2·H2O], NaCl, NaNO3, trichloroacetic
acid (TCA), glacial acetic acid, sodium dihydrogen phosphate, disodium
hydrogen phosphate, anthrone as reagent, Folin-Ciocalteu as reagent,
congo red, cetyltrimethylammonium bromide (CTAB), chitin flakes, Tris–HCl,
β-mercaptoethanol, phenylmethylsulfonyl fluoride (PMSF), and
KCl (all AR grade) were purchased from E. Merck, India. Thymine, cytosine,
and adenine were purchased from TCI, Japan, and calf thymus DNA, crystal
violet (CV), ethyl acetate, hydroxyl amine, NaOH, and ferric chloride
were procured from Sisco Research Laboratories, India. Calf thymus
DNA was dissolved in triple distilled water in the presence of 120
mM NaCl, 35 mM KCl, and 5 mM MgCl2. Its concentration was
determined using a molar extinction coefficient of 6600 M–1 cm–1 at 260 nm. Absorbance of the DNA solution
was also measured at 280 nm; A260/A280 was determined. The value found in the range
1.8–1.9 was considered ready for use, not requiring further
purification. Quality of calf thymus DNA was verified using circular
dichroism (CD), recording its response at 260 nm on a CD spectropolarimeter
(J815—JASCO, Japan). Aqueous solutions of all other substances
were prepared in triple distilled water.
4
Circular Dichroism Analysis of DNA-GSH Interactions
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A Circular Dichroism (CD) Spectropolarimeter (Jasco 815, Easton, MD, United States) was used to investigate DNA-GSH reactions in a 1-mm path length quartz cuvette. dsDNA (calf thymus DNA-sodium salt Type 1 fibers, Sigma-Aldrich, Australia) and GSH stock solutions were prepared in sterile MilliQ water. To study the interaction, 200 ng/μl dsDNA incubated for 24 h at 37°C, 100 rpm, in either absence or presence of 1 mM GSH at intrinsic pH or 1 mM GSH at neutral pH (7.2). 300 μl aliquots pipetted into cuvettes and scanned by CD at 200–320 nm wavelength in a static condition at 25°C.
5
LPS and LTA Binding Assay by CD Spectroscopy
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The LPS and LTA binding assay was performed to investigate the interaction between PN5 and E. coli O111:B4 LPS and S. aureus LTA (Sigma-Aldrich) by CD spectroscopy. E. coli LPS and S. aureus LTA were diluted in 10 mM sodium phosphate buffer (pH 7.4) to 1 mg mL−1. The peptide (50 μM) was added to E. coli LPS and S. aureus LTA. The temperature was regulated by a PTC-423S controller and was set to 20°C. The spectra were measured from 190 to 250 nm using a CD spectropolarimeter (Jasco) operating at room temperature (25°C) (61 (link)).
6
IAPP Structural Analysis in Lipid Membranes
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A freshly prepared stock solution of IAPP (500 μM in water) was diluted to 15 μM and 20 μM in phosphate buffer containing DOPG : DOPC (3 : 7, 750 μM, d = 100 nm) and in phosphate buffer, respectively, for CD measurements. The spectra of IAPP were recorded at 0.5 nm intervals from 190 to 260 nm with an averaging time of 10 s and an average of three repeats on a Jasco CD Spectropolarimeter. Spectra were recorded in presence of DM 1 using the identical method as described above.
7
CD Spectroscopy of Thaumatin-like Protein
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CD spectra were recorded on CD spectropolarimeter (Jasco J‐1500), equipped with a Peltier‐type temperature controller, as previously described (Dindo et al., 2020 (link); Pedretti et al., 2020 (link)). Briefly, near UV–Vis (250–600 nm) spectra of 1 mg/mL TgCGL were collected in 1‐cm path length quartz cuvette at a scan speed of 50 nm/min in 20 mM sodium phosphate pH 8 at 25°C. A minimum of three accumulations were made for each scan, averaged, and corrected for the blank solution of corresponding buffer (Bombardi et al., 2020 (link)).
8
Thermostability Analysis of Laccase
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CD spectra of the purified laccase was recorded at various temperature ranging 30– 65 °C in 0.1 M sodium phosphate buffer (pH 6.8) using a CD Spectropolarimeter (JASCO J– 810) (Yang et al. 1986 (link)).
9
Circular Dichroism Analysis of Peptides
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Circular dichroism
(CD) samples were prepared in PBS or in PBS supplemented with 3 M
KCl or 5 M GuHCl. Samples contained 20 μM peptide. CD spectra
were collected on a JASCO CD Spectropolarimeter over the range of
190–260 nm, and each spectrum was an average of 64 scans. Background
spectra from samples containing no peptide were subtracted from the
sample spectra.
(CD) samples were prepared in PBS or in PBS supplemented with 3 M
KCl or 5 M GuHCl. Samples contained 20 μM peptide. CD spectra
were collected on a JASCO CD Spectropolarimeter over the range of
190–260 nm, and each spectrum was an average of 64 scans. Background
spectra from samples containing no peptide were subtracted from the
sample spectra.
10
Thermal Denaturation Analysis by CD
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