Mice were anesthetized with a mixture of alfaxalone and xylazine (5:1 ratio) and transcardially perfused with cold PBS. Mouse brains were harvested and fixed using 10% NBF (Sigma; HT501128) for 2 h. After washing with PBS for 30 min three times, fixed brains were embedded in OCT compound (Leica; #3801480) for frozen coronal sectioning at a thickness of 7 μm. To detect microglial activation, brain sections were blocked with 5% donkey serum in PBS (Jackson Laboratory; #017-000-121) for 1 h and then incubated with goat anti-Iba-1 antibody (Abcam; ab5076, 1:100 dilution) and rabbit anti-TMEM119 antibody (Abcam; ab209064, 1:100 dilution) at 4 °C overnight. The sections were visualized with Alexa Fluor 488-conjugated donkey anti-goat IgG antibody (Jackson Laboratory; #705-545-003, 1:200 dilution) and Alexa Fluor 568-conjugated donkey anti-rabbit IgG (Abcam; ab175693, 1:300 dilution). Confocal microscopy (Zeiss; LSM800) in the Korea Mouse Phenotyping Center (KMPC) was used to analyze the Iba-1-positive area of sections.
Alexa fluor 488 conjugated donkey anti goat igg antibody
Manufactured by Jackson ImmunoResearch
Alexa Fluor 488-conjugated donkey anti-goat IgG antibody is a secondary antibody that binds to goat immunoglobulin G (IgG) and is conjugated to the Alexa Fluor 488 fluorescent dye. It is used for the detection and visualization of goat primary antibodies in various immunoassays and imaging applications.
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2 protocols using alexa fluor 488 conjugated donkey anti goat igg antibody
1
Microglial Activation Analysis in Mouse Brains
2
Immunofluorescent Localization of D2R and KOR
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C57BL/6 mice were anesthetized with a mixture of ketamine/xylazine (50/5 mg/kg, respectively) and fixed by cardiac perfusion with 2% paraformaldehyde. Immunofluorescence studies in brain slices of 40 µm were performed with the following antibodies: goat polyclonal anti-D2R antibody (1:250, sc-7522, Santa Cruz Biotechnology) and rabbit polyclonal anti-KOR antibody (1:1000, Ab10566, Abcam), followed by AlexaFluor 488-conjugated donkey anti-goat IgG antibody (1:1000, Jackson Immunoresearch) and CY3-conjugated donkey anti-rabbit IgG antibody (1:1000, Jackson Immunoresearch). Fluorescence imaging was acquired on a laser-scanning confocal microscope (Olympus, Fluoview 1000) coupled to Fluoview v6.0 software Olympus. Images were obtained with a 100× oil immersion objective.
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