Imaging and uncaging were performed using a two-photon laser scanning microscope (MOM; Sutter). The light source for fluorescence excitation (800 nm for Alexa Fluor 594 and 940 nm for gephyrin intrabody) and RuBi-GABA uncaging (800 nm) was a Ti:Sapphire laser (Chameleon XR; Coherent). LiGABAR-expressing hippocampal neurons were voltage-clamped at 0 mV, with 25 μM DNQX, 50 μM D-AP5, and 0.5 μM TTX in the bath. Internal solution included 200 μM Alexa Fluor 594 (Life Technologies) for visualizing dendritic morphology. RuBi-GABA (200-400 μM; Abcam) was added to aCSF and re-circulated using a peristaltic pump (Idex). Uncaging was carried out at designated locations for 5-10 ms with a light intensity of ~150 mW. Full-field 390 nm (1.2 mW/mm2) or 540 nm (3.2 mW/mm2) conditioning flashes (5 sec) from an LED light source (Lumencor) were delivered through the objective. Photoswitching was calculated as 1 - (I540/I390), where I refers to the peak amplitude of GABA-elicited current.
Rubi gaba
Manufactured by Abcam
Sourced in United Kingdom
RuBi-GABA is a caged GABA compound that can be uncaged with light. It is a biologically active molecule that can be used to study GABA-mediated neurotransmission and related processes.
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Lab products found in correlation
3 protocols using rubi gaba
1
Photoswitchable GABA Uncaging in Neurons
2
Sprague-Dawley Rat Experiment with RuBi-GABA
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Adult male Sprague-Dawley rats weighing 250–350 g were used in our experiment and were acquired from the animal facility of Capital Medical University. Animal care and protocols were approved by the Ethics Committee on Animals Care and Usage of Capital Medical University. RuBi-GABA was purchased from Abcam Biochemicals (Cambridge, UK). Isoflurane was purchased from Ruiwode (Shenzhen, China), and all other chemicals were purchased from Sigma-Aldrich (St Louis, MO, USA).
3
RuBi-GABA Uncaging Experiments
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RuBi-GABA [5μM] for uncaging experiments was purchased from Abcam (Cambridge, UK) and stock solutions were made and diluted within one week of use. Wide-field, 50 ms, 500 ms, and 1 min pulses from a CooLED (Andover, UK) pe-100 LED centered at 470nm (100% power) were used to uncage RuBi-GABA around the soma of the recorded cell.
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