To record the CYP125A13 electron paramagnetic resonance (EPR) spectra, a Bruker X-band spectrometer (9.5 GHz) equipped with a continuous supply of helium cryostat ESR 900 and an ITC 4 temperature controller used to achieve a base temperature of 5 K, was used. The microwave frequency and modulation frequency used were 9.647GHz and 100 kHz, respectively. The data were recorded and analyzed at The Korea Basic Science Research Institute, Western Seoul Center. Before the measurement, the oxidized form of 180 μM CYP12A13 in 10 mM potassium phosphate buffer (pH 7.4) was frozen in liquid nitrogen in EPR quartz tubes (Wilmad). The spectral data comprising the g-tensor and line width of the heme centers were acquired by the accumulation of spectra in the Simfonia or Xsophe (Bruker) programs.
Xsophe
Manufactured by Bruker
Xsophe is a versatile software package developed by Bruker for the analysis and simulation of electron paramagnetic resonance (EPR) spectra. It provides a comprehensive set of tools for the interpretation and modeling of experimental EPR data.
Automatically generated - may contain errors
Lab products found in correlation
Simfonia, by Bruker (1 mentions)
Esr 900, by Bruker (1 mentions)
Epr quartz tubes, by Wilmad (1 mentions)
Elexsys 500 spectrometer, by Bruker (1 mentions)
Esr900 helium flow cryostat, by Oxford Instruments (1 mentions)
Emxmicro spectrometer, by Bruker (1 mentions)
E580 spectrometer, by Bruker (1 mentions)
4 protocols using xsophe
1
Characterization of CYP125A13 by EPR
2
X-band EPR Spectroscopy Protocol
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X-band EPR spectra were recorded on a Bruker ELEXSYS 500 spectrometer equipped with a Bruker ER 4116DM X-band resonator, an Oxford Instrument continuous flow ESR 900 cryostat, and an Oxford ITC 503 temperature control system.
Conditions: Microwave frequency = 9.636 GHz, microwave power = 1.0 milliwatt, modulation amplitude = 8 Gauss, modulation frequency = 100 KHz, Gain = 40 db, temperature =10 K.
Simulations were done using the Bruker software XSophe.
Conditions: Microwave frequency = 9.636 GHz, microwave power = 1.0 milliwatt, modulation amplitude = 8 Gauss, modulation frequency = 100 KHz, Gain = 40 db, temperature =10 K.
Simulations were done using the Bruker software XSophe.
3
EPR Spectroscopy of Cobalt Complexes
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Low temperature
EPR spectra were collected with a Bruker Elexsys E600 EPR spectrometer
equipped with an Oxford Instruments ESR900 helium flow cryostat. Spectra
were recorded at 9.63 GHz (B0⊥B1) or
9.37 GHz (B0∥B1) using an ER4116DM dual-mode
cavity, with 10 G (1 mT) magnetic field modulation at 100 kHz. Microwave
powers and temperatures employed are given in the legends to figures,
and other recording parameters were chosen such that resolution was
limited by the field modulation amplitude. Spin Hamiltonian parameters
were estimated from computer simulations carried out using XSophe
(Bruker Biospin), assuming Ĥ = βgBS + SDS + ASI, where S = 3/2, D ≫ βgBS, and I(59Co) = 7/2, and where D > 0 implies MS = ± 1/2 and D < 0 implies MS = ± 3/2.28
EPR spectra were collected with a Bruker Elexsys E600 EPR spectrometer
equipped with an Oxford Instruments ESR900 helium flow cryostat. Spectra
were recorded at 9.63 GHz (B0⊥B1) or
9.37 GHz (B0∥B1) using an ER4116DM dual-mode
cavity, with 10 G (1 mT) magnetic field modulation at 100 kHz. Microwave
powers and temperatures employed are given in the legends to figures,
and other recording parameters were chosen such that resolution was
limited by the field modulation amplitude. Spin Hamiltonian parameters
were estimated from computer simulations carried out using XSophe
(Bruker Biospin), assuming Ĥ = βgBS + SDS + ASI, where S = 3/2, D ≫ βgBS, and I(59Co) = 7/2, and where D > 0 implies MS = ± 1/2 and D < 0 implies MS = ± 3/2.28
4
Magnetic Susceptibility and EPR Characterization
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Variable-temperature
(2–290
K) magnetic susceptibility data were recorded in 1 T external field
using an MPMS Quantum Design SQUID magnetometer. The experimental
data were corrected for underlying diamagnetism using tabulated Pascal’s
constants. S- and X-band fluid solution spectra were collected using
a Bruker EMX Micro spectrometer, and frozen solution spectra were
obtained using a Bruker E580 spectrometer. Simulations were performed
using the Xsophe (Bruker Biospin GmbH) suite.44 (link) The elemental analysis was performed by Midwest Microlab, LLC, of
Indianapolis, IN.
(2–290
K) magnetic susceptibility data were recorded in 1 T external field
using an MPMS Quantum Design SQUID magnetometer. The experimental
data were corrected for underlying diamagnetism using tabulated Pascal’s
constants. S- and X-band fluid solution spectra were collected using
a Bruker EMX Micro spectrometer, and frozen solution spectra were
obtained using a Bruker E580 spectrometer. Simulations were performed
using the Xsophe (Bruker Biospin GmbH) suite.44 (link) The elemental analysis was performed by Midwest Microlab, LLC, of
Indianapolis, IN.
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