All sections received CB-HRP immunofluorescent staining (rabbit anti-CB primary antibody diluted in 1:600, Abcam; donkey anti-rabbit Alexa Fluor 488 secondary antibody diluted in 1:200, Life Technologies). Then, the sections were mounted in sequence on the slides, counterstained by DAPI, and coverslipped. The sections of different positions were captured at uniform parameter using a fluorescence microscope (Leica DM6, Germany). The number of positive neurons was counted by the Image-Pro Plus 7.0 software. Part of the sections received CB-HRP/serotonin immunofluorescent double labeling to confirm the neurotransmitter (rabbit anti-CB primary antibody diluted in 1:600, Abcam; goat anti-serotonin primary antibody diluted in 1:600, Abcam; donkey anti-rabbit Alexa Fluor 488 secondary antibody diluted in 1:200, Life Technologies; donkey anti-goat Alexa Fluor 594 secondary antibody diluted in 1:200, Life Technologies).
Rabbit anti cb primary antibody
Manufactured by Abcam
Sourced in United States
Rabbit anti-CB primary antibody is a laboratory reagent used to detect the presence of CB proteins in biological samples. It is a polyclonal antibody produced in rabbits and specifically targets CB proteins.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 donkey anti rabbit secondary antibody, by Thermo Fisher Scientific (3 mentions)
Donkey anti goat alexa fluor 594 secondary antibody, by Thermo Fisher Scientific (1 mentions)
Lsm 880, by Zeiss (1 mentions)
Laser confocal microscope, by Zeiss (1 mentions)
Alexa fluor 488 secondary antibody, by Thermo Fisher Scientific (1 mentions)
4 protocols using rabbit anti cb primary antibody
1
Calretinin-Containing Neuron Visualization
2
Immunofluorescent Staining and Quantification of Calretinin-Positive Neurons
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After CB immunofluorescent staining (rabbit anti-CB primary antibody diluted in 1:600, Abcam; donkey anti-rabbit Alexa Fluor 488 secondary antibody diluted in 1:200, Life Technologies), the sections were sequentially mounted on slides, counterstained with DAPI, and coverslipped. The sections of the brainstem and spinal cord were visualized using fluorescence (Leica DM6, Germany) and confocal laser microscopes (Zeiss LSM 880, Germany) for different positions at the uniform standard. The cell density of CB-positive neurons (cell number/0.2 mm2 area) in each brain region was calculated using Image-Pro Plus 7.0 software. The density of CB-positive neurons was classified according to the densities: <5, 6–10, and >10, which correspond to sparse, moderate, and dense distributions, respectively.
3
Immunofluorescence Analysis of Calbindin-Positive Neurons
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All sections were examined with CB immunofluorescence (rabbit anti-CB primary antibody diluted in 1:600, Abcam; donkey anti-rabbit Alexa Fluor 488 secondary antibody diluted in 1:200, Life Technologies, Carlsbad, CA, USA). Sections were mounted in sequence on slides, counterstained with DAPI and coverslipped. Diencephalon sections were imaged with a fluorescence microscope (Leica DM6, Germany) and a confocal laser microscope (Zeiss, Germany). The cell density of CB-positive neurons (cell number/0.2 mm2 area) in each brain region was calculated using Image-Pro Plus 7.0 software. The density of CB-positive neurons was classified as sparse, moderate and dense according to the densities: <5, 6–10 and >10, respectively.
4
Immunofluorescence Staining of Calbindin
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All the sections were subjected to CB immunofluorescence staining. The antisera used for immunofluorescence processing were diluted in a solution of 0.1 M PBS containing 0.3% Triton X-100. The sections were incubated in rabbit anti-CB primary antibody (1:600, Cat# ab34992, Abcam) for 48 h at 4°C. Next, the slices were incubated in donkey anti-rabbit Alexa Fluor 488 secondary antibody (1:200, Cat# A-21206, Thermo Fisher) for 2 h in the dark at room temperature. Then, the sections were mounted in sequence onto slides, counterstained with 4’,6-diamidino-2-phenylindole (DAPI), and covered with coverslips. Images of the sections of the subcortex and limbic system were captured under a fluorescence microscope (DM6, Leica, Germany) and confocal laser microscope (Zeiss, Germany). The 10× lens was used to capture the positive neurons, while the 40× lens was applied to show the detailed structures.
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