To follow chemical induced unfolding, we prepared samples with 1.4 μM protein concentration in phosphate buffer (described earlier), containing different concentrations of GuHCl/urea. Samples were incubated for 12 hours to reach equilibrium at 25°C, after which no change in signal occurred either in fluorescence or CD spectra. Trp fluorescence (excitation at 295 nm and emission recorded between 300 nm to 400 nm) and ANS fluorescence (excitation at 370 nm and emission recorded between 400 nm to 600 nm) measurements were performed using a Hitachi F-7000 fluorescence spectrophotometer for GuHCl samples and Jasco J-815 spectropolarimeter for urea samples. Far-UV CD measurements were performed using Jasco J-810 spectropolarimeter for GuHCl samples and Jasco J-815 spectropolarimeter for urea samples. Quartz cuvette of 1 cm and 0.5 cm path-length were used for fluorescence and CD measurements, respectively. All measurements were done at 25 °C. We have reported spectra as ellipticity (millidegree) after baseline correction.
J 815 spectropolarimeter
Manufactured by Jasco
Sourced in Japan, United States, Germany, United Kingdom, Italy, Canada
The J-815 spectropolarimeter is a laboratory instrument used for the measurement of circular dichroism (CD) spectra. It is designed to analyze the interaction between polarized light and chiral molecules or structures. The J-815 spectropolarimeter provides accurate and reliable data on the secondary structure and conformational changes of biological macromolecules such as proteins, nucleic acids, and other chiral compounds.
Automatically generated - may contain errors
Lab products found in correlation
Sephadex lh 20, by GE Healthcare (10 mentions)
Cdf 426s 15, by Jasco (7 mentions)
Silica gel, by Qingdao Marine Chemical (6 mentions)
Acquity uplc beh c18 column, by Waters Corporation (6 mentions)
Silica gel, by Qingdao Haiyang Chemical (6 mentions)
Avance 3 500 spectrometer, by Bruker (5 mentions)
Peltier temperature controller, by Jasco (5 mentions)
Sephadex lh 20, by Cytiva (5 mentions)
Uv 2550, by Shimadzu (4 mentions)
Analytical automatic polarimeter, by Shimadzu (4 mentions)
Spectra manager software, by Jasco (4 mentions)
Ptc 423, by Jasco (4 mentions)
Vertex 70v ftir spectrometer, by Bruker (4 mentions)
Avance 500 spectrometer, by Bruker (4 mentions)
Ptc 423s 15, by Jasco (4 mentions)
Pfd 425s, by Jasco (4 mentions)
Agilent 500 mhz dd2 spectrometer, by Agilent Technologies (4 mentions)
Spectra manager 2, by Jasco (3 mentions)
Arx 600 spectrometer, by Bruker (3 mentions)
Ltq orbitrap xl spectrometer, by Thermo Fisher Scientific (3 mentions)
628 protocols using j 815 spectropolarimeter
1
Protein Unfolding Kinetics by Spectroscopy
2
Circular Dichroism Spectroscopy of hTAS1R2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The circular dichroism (CD) spectra of the FLAG-tagged hTAS1R2 samples were recorded at 20 °C using a J-815 Jasco spectropolarimeter (Jasco, Tokyo, Japan) equipped with a Peltier temperature control. The CD spectra were corrected for the buffer and ligand contributions and converted to mean residue ellipticity in deg cm2 dmol−1. The spectra recorded between 178 and 260 nm were averaged over 5 scans accumulated at 0.5 nm intervals with a 50 nm/min scan speed and 5 s of response time. Spectra were smoothed using the Savitzky-Golay convolution filter with a span of 15. The secondary structure proportions were estimated using the deconvolution CDSSTR algorithm available on the DichroWeb website (http://dichroweb.cryst.bbk.ac.uk/html/home.shtml )73 (link).
3
Circular Dichroism Analysis of Histone H1 Structures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Circular Dichroism evaluates the changes in protein secondary structure, folding and their binding properties. We carried out the Far-UV CD spectral analysis of native histone H1 and its MG modified counterpart using J-815 JASCO spectropolarimeter. The instrument was attached to a microcomputer with a thermostatically controlled cell holder attached to Neslab's RTE 110 water bath with a temperature accuracy of ±0.1°C. The scans were taken at wavelength intervals of 1 nm at 25°C. All scans were recorded at wavelength intervals of 1 nm. Spectra were collected at 50 nm min-1 scan speeds, 0.1 nm data pitch and a response time of 2s. The spectra were recorded in far UV region between wavelengths 190 to 250 nm and the protein concentration was 0.2 mg/ml. The results were obtained in molar residual ellipticity (θ) (deg/cm2/dmol) at a wavelength λ, based on the equation given below [24 ].
Where θobs is the observed ellipticity in degrees, Cp is the molar fraction and l is the length of the light path in centimeter. The α-helical content of different histone H1 samples were calculated from θ values at 222 nm (MRE 222) using the following equation.
Where θobs is the observed ellipticity in degrees, Cp is the molar fraction and l is the length of the light path in centimeter. The α-helical content of different histone H1 samples were calculated from θ values at 222 nm (MRE 222) using the following equation.
4
Cationic Surfactant-Nucleic Acid Binding Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine whether structural changes in DNA or RNA occurred upon binding of cationic surfactants to nucleic acids, circular dichroism spectra were recorded on a J-815 JASCO spectropolarimeter. All CD measurements were carried out at room temperature (25 ± 1°C) using a 1 mm quartz cuvette. Scans were obtained in the UV range between 220 and 350 nm, at a scan rate of 100 nm/min with bandwidth of 1 nm and integration time of 2 s. Each time, five subsequent spectra were averaged to improve the signal-to-noise ratio. The background CD spectrum recorded for the buffer solution was subtracted from the samples spectra, which were then smoothed using the Savitzky-Golay function over 9 points. All of the data were analysed using Spectra Manager II (Jasco) and Origin software.
5
Circular Dichroism Analysis of IgG1-Fc
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The secondary structure of the IgG1-Fc domian (from P. pastoris WT and MNN9-EndoT strains) were determined by circular dichroism using J-815 Jasco spectropolarimeter (Jasco Co., Tokyo, Japan) equipped with a PTC-348 WI thermostat under a constant nitrogen flow. A 0.1-cm path length cell was used to collect data in the far ultraviolet region (200–250 nm) at a scan speed of 20 nm/min and a response time of 1 s. Spectra were acquired at 25 °C and measured in PBS buffer. The spectrum of a blank containing buffer alone was subtracted from all spectra. The CD data were analyzed using the CDtoolX and online tools dichroweb (http://dichroweb.cryst.bbk.ac.uk/ ).
6
Far-UV CD Spectroscopy of Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD spectra were obtained in the far-UV (260–195 nm) on a J-815 Jasco spectropolarimeter (Tokyo, Japan) at 37 °C, using a 10 mm path cell, with 2 accumulations for a 216.0 μg/mL sample in 10 mM sodium phosphate buffer, pH 6.6.
7
Sponge Extract Secondary Structure Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD spectroscopy was applied to assess alterations in the secondary structure of the Col molecule in the marine sponge extracts. Spectra of extracts in the concentration of 1 mg/mL were recorded with a J-815-JASCO® spectropolarimeter (JASCO®, São Paulo, Brazil) in 0.5 mm path length quartz cuvettes from 190–250 nm at 25 °C, with a scan speed of 50 nm/s. The final spectra obtained was the average of four consecutive measurements. Distilled water was used as a blank.
8
Structural Changes of DNA in Surfactants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CD spectra were recorded to investigate structural changes of DNA occurred after adding surfactants. A J-815 JASCO spectropolarimeter and a 1 mm quartz cuvette were used. Spectra were recorded in the UV range (220–350 nm), at a scan rate of 100 nm/min with bandwidth of 1 nm and integration time of 2 s. To achieve a better signal-to-noise ratio, each spectrum was calculated as the average of five recorded scans. All measurements were carried out at room temperature (25 ± 1 °C). A background CD spectrum recorded a 5 mM phosphate buffer, which was subtracted from the sample spectra. All spectra were smoothed using the Savitzky-Golay function over 9 points. For analysis and presentation of obtained data, Spectra Manager II (Jasco, Tokyo, Japan) and Origin software were used. Due to high turbidity of samples with phospholipids, measurements were not possible.
9
Characterization of Organic Compounds
10
Comprehensive Analytical Techniques for Chemical Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Optical rotations were recorded on an Autopol IV automatic polarimeter. The CD spectra were measured on a JASCO J-815 spectropolarimeter. UV spectra were obtained on a Persee TU-1901 UV-vis spectrometer. 1D and 2D NMR spectra were performed at 600 MHz for 1H NMR and 150 MHz for 13C NMR on Bruker ARX-600 spectrometer. Chemical shifts (δ) are given in ppm, and coupling constants (J) are given in hertz (Hz). ESIMS data were recorded on a Thermo LTQ mass spectrometer. HRESIMS data were measured using a Thermo LTQ Orbitrap XL mass spectrometer. Column chromatography (CC) were carried out with silica gel (200–300 mesh, Qingdao Marine Chemical Inc. Qingdao, PR China). Analytical TLC was carried out on pre-coated silica gel GF254 plates (Qingdao Marine Chemical Industry, Qingdao, China), and spots were visualized under UV light.
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
Revolutionizing how scientists
search and build protocols!