The largest database of trusted experimental protocols

Chirascan cd spectrometer

Manufactured by Applied Photophysics
Sourced in United Kingdom

The Chirascan CD spectrometer is a laboratory instrument designed to measure the circular dichroism (CD) of samples. CD is a spectroscopic technique that provides information about the structural properties of chiral molecules, such as proteins and other biomolecules. The Chirascan CD spectrometer is capable of accurately measuring CD signals over a wide range of wavelengths, enabling researchers to study the conformation and folding of these molecules.

Automatically generated - may contain errors

295 protocols using chirascan cd spectrometer

1

Amyloid Aggregation Kinetics Modulated by ATP

Check if the same lab product or an alternative is used in the 5 most similar protocols
Monomeric NM (2.5 μM) was aggregated in assembly buffer (40 mM Hepes–KOH [pH 7.4], 150 mM KCl, 1 mM DTT, 20 mM MgCl2) without or with several concentrations of ATP (2.5 nM, 2.5 μM, 2.5 mM, and 10 mM) at room temperature with stirring at 80 rpm for 6 h. The amyloid species were pelleted after 6 h at 16,400 rpm for 30 min and resuspended in 10 mM Na2PO4 (pH 7.4) buffer. The final concentration of NM was ∼8 μM, after which far-UV CD spectra were recorded on a Chirascan CD spectrometer (Applied Photophysics) at room temperature. All the spectra were collected and recorded in the scan range 195 to 260 nm with a 1 nm step size and a quartz cuvette of 1 mm path length. The spectra were averaged over 5 to 10 scans, and the buffer signal was subtracted. All the spectra were smoothened using the ProData software (Applied Photophysics) provided with the Chirascan CD spectrometer. Finally, the mean residue ellipticity [θ] was calculated, and plots were generated using Origin software.
+ Open protocol
+ Expand
2

Structural Analysis of Rpgrip1l Domains

Check if the same lab product or an alternative is used in the 5 most similar protocols
CD spectra of mouse Rpgrip1l CC12, CC1, CC2, CC12 and human Rpgrip1 CC12 wild-type and mutations at a concentration of 0.2 mg/ml in buffer containing 50 mM Tris-HCl, pH 7.5, 1 mM EDTA, 1 mM DTT, 250 mM Na2SO4 were acquired on a Chirascan CD Spectrometer (Applied Photophysics) at 25 °C using a cell path length of 1 mm. Each spectrum was collected with three scans spanning a spectral window of 200∼280 nm. Thermal denaturation experiments were performed on the Chirascan CD Spectrometer (Applied Photophysics) from 20∼95 °C at a 1 °C stepped temperature ramping with a setting time of 30 sec. In both denaturation experiments, the ellipticities at 222 nm were used to calculate the unfolded fraction and plotted as a function of temperature.
+ Open protocol
+ Expand
3

Quantifying Membrane Alignment via Linear Dichroism

Check if the same lab product or an alternative is used in the 5 most similar protocols
Linear dichroism (LD) spectra were recorded on a Chirascan CD spectrometer between 200 and 350 nm in 1 nm increments at a time per point of 0.7 s and a bandwidth of 1 nm. The alignment of the liposomes was achieved by a custom-made outer-cylinder-rotation Couette flow cell with a path length of 1 mm. The shear rate was 3100 s−1. Baselines at zero shear gradients were collected and subtracted from all spectra. The macroscopic orientation of the membrane was probed using curcumin absorption band at 424 nm (Rocha et al., 2016 (link)). Stock solutions of curcumin (Sigma-Aldrich) were prepared with ethanol (99.7%), and a specific volume (less than 0.5% of the total sample volume) was added to the vesicles in order to have a final concentration of about 2 μM. The measurements were done in a high viscosity buffer (containing 50 wt% sucrose) to reduce the light scattering of the liposomes by matching their refractive index (Ardhammar et al., 2002 (link); Rocha et al., 2016 (link)).
+ Open protocol
+ Expand
4

Far UV-CD Analysis of Protein Structure

Check if the same lab product or an alternative is used in the 5 most similar protocols
Far UV-CD analysis was performed using a Chirascan™ CD spectrometer. Data was recorded over a wavelength range of 190–260 nm with a 0.5 nm step size. 1 mg ml–1 samples in 100 mM ammonium acetate buffer (concentration measured precisely using a Thermo NanoDrop 2000) were added to a 0.1 mm cuvette and analysed at 25 °C for 12 minutes (time per point = 5 s). The data was converted from millidegrees (mdeg) to molar ellipticity ([θ]) to correct for deviations in concentration between samples.
+ Open protocol
+ Expand
5

Spectroscopic Analysis of Natural Compounds

Check if the same lab product or an alternative is used in the 5 most similar protocols
Similar to previous procedures [10 (link),15 (link),16 (link),17 (link),23 (link)], Autopol VI polarimeters were applied to determine optical rotations. The Chirascan CD spectrometer was applied to measure UV and ECD data. The Nicolet iS50 FT-IR spectrometer was applied to acquire IR data. The Bruker Avance III 500 NMR spectrometer (500 MHz for 1H and 125 MHz for 13C) was applied to record 1D/2D NMR data. The Xevo G2-XS QTof mass spectrometer was applied to obtain HRESIMS data. The Agilent 1260 Infinity II system with a ZORBAX SB-C18 (5 μm, 9.4 × 250 mm) column was applied for HPLC separation. Silica gel (200–300 mesh, Qingdao Haiyang Chemical Co., Qingdao, China), RP-18 (AAG12S50, YMC Co., Ltd., Kyoto, Japan), and Sephadex LH-20 (GE Healthcare, Chicago, IL, USA) were employed for column chromatography (CC). Precoated silica gel plates (GF-254, 20 × 20 cm, Qingdao Haiyang Chemical Co., Qingdao, China) were used for thin-layer chromatography (TLC).
+ Open protocol
+ Expand
6

Spectroscopic Characterization of Compounds

Check if the same lab product or an alternative is used in the 5 most similar protocols
Chemical shifts are reported in parts per million from TMS. All NMR spectra were recorded on an Agilent 400-MR-NMR spectrometer operated at 400 and 100 MHz for hydrogen and carbon, respectively. Data processing was carried out with the MestReNova ver.6.0.2 program. The IR spectra were obtained from a Tensor 37 FT-IR spectrometer (Bruker, Ettlingen, Germany). UV and Circular dichroism spectrums were determined on a Chirascan™ CD spectrometer. Preparative HPLC was carried out using an AGILENT 1200 HPLC system. Column chromatography (CC) was performed on silica-gel (Kieselgel 60, 70–230 mesh and 230–400 mesh, Merck, Kenilworth, NJ, USA), YMC RP-18 resins (30–50 μm, Fujisilisa Chemical Ltd., Kasugai, Aichi, Japan), or Diaion HP-20 regins (200–300 mesh, Mitsubishi Chemical Co., Chiyoda, Tokyo, Japan). For thin-layer chromatography (TLC), pre-coated silica-gel 60 F254 (0.25 mm, Merck) and RP-18 F254S (0.25 mm, Merck) plates were used.
+ Open protocol
+ Expand
7

CD Analysis of Peptide Release

Check if the same lab product or an alternative is used in the 5 most similar protocols
To analyse the release of peptides from fibres into solution, CD analysis of the supernatant fluid was conducted and spectra were recorded using a Chirascan CD spectrometer with 1 mm path-length cuvettes at 22 °C. The data were acquired at a step resolution of 1 nm and scan speed of 60 nm min−1. A bandwidth of 4.3 nm was used to obtain smoother spectra. Far-UV spectra were recorded in the wavelength range 185 to 260 nm. Each spectrum was the average of two scans and the spectrum for the blank solvent was subtracted. The data then were converted to mean residue ellipticity (deg cm2 dmol−1) and fitted with a polynomial equation (R2 ≥ 0.95).
+ Open protocol
+ Expand
8

Quantifying Fibrillar α-Synuclein using UV-CD

Check if the same lab product or an alternative is used in the 5 most similar protocols
A Chirascan CD
spectrometer was used to obtain UV-CD spectra of αS fibrils
prepared in PBS buffer at an effective protein concentration of 10
μM. This was measured as follows: after measurement of fibril
UV-CD spectra, 30% 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) was added
to the fibril suspension and shaken for 5 min. By measuring mean residue
ellipticity (MRE) values for known concentrations of monomeric αS
obtained after addition of 30% HFIP and shaking, a standard curve
of UV-CD signals versus monomer concentration was generated. From
this curve, unknown concentrations of 30% HFIP-solubilized fibrils
were estimated. Fibril samples were first purified using a 100 kDa
cutoff filter to remove monomeric protein. Spectra were recorded between
195 and 260 nm with a step size of 1 nm and a scanning speed of 10
nm/min using a 1 mm path length cuvette at room temperature.
+ Open protocol
+ Expand
9

Spectroscopic Characterization of Compounds

Check if the same lab product or an alternative is used in the 5 most similar protocols
Optical rotations were recorded on a Polax-2L polarimeter in MeOH. HR-ESIMS were recorded on a FT-ICR 910-MS TQFTMS-7 T mass spectrometer. CD spectra were taken on a Chirascan CD spectrometer. IR spectra were recorded on a Nicolet Impact 410 FT-IR spectrometer, and NMR spectra on a Bruker AM500 MHz spectrometer operating at 125.76 MHz for 13C NMR, and at 500.13 MHz for 1H NMR. 1H chemical shifts were referenced to CD3OD at δ 3.31 ppm, while the 13C chemical shifts were referenced to the central peak at δ 49 ppm. For HMBC experiments the delay (1/2J) was 70 ms.
+ Open protocol
+ Expand
10

Spectroscopic Characterization of Compounds

Check if the same lab product or an alternative is used in the 5 most similar protocols
NMR spectra were recorded on a 500 MHz Bruker AVANCE NMR spectrometer in either CDCl3 or CD3OD, UV-VIS spectra were recorded on a Libra Biochrom spectrometer in CH3OH in a 1 cm cell, IR spectra were obtained using an Agilent (Cary 600 series) FTIR spectrometer (University of Surrey), ESIMS analysis was performed using an Alliance 2695 Quattro Ultra mass spectrometer, HRESIMS data were recorded on an Agilent 6550 iFunnel Q-TOF LC/MS with samples dissolved in CH3OH. Optical rotations were measured at room temperature in CH3OH using a JASCO P-1020 polarimeter and CD spectra were measured on a Chirascan CD spectrometer using a 1 mm cell in CH3CN. Solvents were reagent grade and purchased from Sigma-Aldrich.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!