At the end of each indicated treatment, 0.5 ml CM was collected for NO detection with EPR as described previously [44 (link), 45 (link)]. In brief, the NO spin trap Fe(MGD)2 was freshly prepared by mixing a stock solution of ferrous sulfate and NaMGD (Enzo) in a molar ratio of 1:5 in distilled water, anaerobically, prior to each experiment. Right after the addition of 1 mM Fe(MGD)2, bEND3 cells (2×106) were exposed to OGD or normoxia for 2 h at 37 °C with or without the presence of the iNOS specific inhibitor 1400W (1 µM, Cayman). At the end of exposure, 0.5 ml CM were immediately transferred into custom-made gas permeable Teflon tubing (Zeus Industries), folded four times, and inserted into a quartz EPR tube for measuring NO spectrum using Bruker EleXsys 540 X-band EPR spectrometer (Billerica, MA) operating at 9.03 GHz and 100 kHz field modulation and spectra. Instrument settings were as follows: magnetic field, 3443 G; scan range, 100 G; microwave power, 21 mW; modulation frequency, 100 kHz; modulation amplitude, 1.0 G; and time constant, 20 ms. The EPR spectra were collected, stored, and manipulated using the Bruker Software Xepr (Billerica, MA).
Elexsys 540 x band epr spectrometer
Manufactured by Bruker
The EleXsys 540 X-band EPR spectrometer is a laboratory equipment designed for electron paramagnetic resonance (EPR) spectroscopy. It operates at X-band microwave frequencies to detect and analyze paramagnetic species in a sample. The instrument provides the core functionality for performing EPR measurements and data acquisition.
Automatically generated - may contain errors
2 protocols using elexsys 540 x band epr spectrometer
1
Nitric Oxide Detection in Hypoxia-Exposed Cells
2
ATP and ROS Quantification in Transgenic Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ATP levels in brains of Tg mice were determined using an ATP Bioluminesence Assay Kit (Roche) following the manufacturer's instruction. Brain tissues were homogenized in the lysis buffer provided in the kit, incubated on ice for 15 min, and centrifuged at 14,000 g for 15 min. Subsequent supernatants were measured for the ATP levels using Luminescence plate reader (Molecular Devices) with an integration time of 10 seconds.
Evaluation of intracellular ROS levels was accessed by election paramagnetic resonance (EPR) spectroscopy as described in our previous study (Du et al.,2017 ). CMH (cyclic hydroxylamine 1‐hydroxy‐3‐methoxycarbonyl‐2,2,5,5‐tetramethyl‐pyrrolidine, 100 μM) was incubated with hippocampal slices for 30 min and then washed with cold PBS. The tissues were collected and homogenized with 100 μl of PBS for EPR measurement. The EPR spectra were collected, stored, and analyzed with a Bruker EleXsys 540x‐band EPR spectrometer (Billerica, MA) using the Bruker Software Xepr (Billerica, MA).
Evaluation of intracellular ROS levels was accessed by election paramagnetic resonance (EPR) spectroscopy as described in our previous study (Du et al.,
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!