For FGF2 immunostaining, brain sections were prepared as described 34 (link). The brains of 12 NF-α1-KO mice and 12 WT littermates at 6-, 14-, and 30-wk of age (2 male and 2 female mice of each genotype, sex, and age) were dissected from perfused mice and embedded (24 mouse brains per block) in a gelatin matrix using MultiBrain™ Technology (NeuroScience Associates, Knoxville, TN). The MultiBrain™ block was sectioned in the coronal plane at 35 μm on an AO 860 sliding microtome. Every 8th free-floating section was used for FGF2 immunohistochemistry and was stained as described 34 (link), using a rabbit anti-FGF2 antibody (1:1,500 dilution, LifeSpan Biosciences). For DCX staining, 35 μm brain sections from perfused FGF2 treated or naïve WT and CPE-KO mice were cut under contract by NeuroScience Associates. The sections were then stained for DCX using polyclonal rabbit anti-doublecortin (1:2,000, Cell Signaling, Danvers, MA) followed by Alexa Fluor 594 goat anti-rabbit secondary antibody (Invitrogen, Carlsbad, CA). Every 10th free-floating section was used for DCX immunostaining.
Polyclonal rabbit anti doublecortin
Manufactured by Cell Signaling Technology
Sourced in United States
Polyclonal rabbit anti-doublecortin is a primary antibody that binds to the doublecortin protein. Doublecortin is a microtubule-associated protein involved in the development and migration of neurons.
Automatically generated - may contain errors
3 protocols using polyclonal rabbit anti doublecortin
1
FGF2 and DCX Immunostaining in Mouse Brain
2
Analyzing Alzheimer's Pathology in Transgenic Mice
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The brains of 50-week-old WT and CPE-QQ transgenic mice were dissected from perfused mice and embedded in a gelatin matrix using MultiBrain Technology (NeuroScience Associates, Knoxville, TN, USA) or each brain was embedded and sectioned, all the sections were coronal. The MultiBrain block was sectioned in the coronal plane at 35 μm on an AO 860 sliding microtome. The sections were then stained for doublecortin (DCX), p-tau, or MAP2 using polyclonal rabbit anti-doublecortin (1:2000, Cell Signaling), polyclonal rabbit anti-p-tauSer396 (1:1000, Santa Cruz), and monoclonal mouse anti MAP2 antibody (1:3000, Abcam, Cambridge, MA, USA), respectively, followed by Alexa Fluor 594 goat anti-rabbit secondary antibody (Invitrogen). After mounting of the sections, images were taken from the hippocampus by a confocal microscope (Zeiss LSM 510 Inverted Meta, Carl Zeiss Microscopy, Thornwood, NY, USA) with × 20 or × 63 magnification. For counting of DCX immunoreactive cells in the dentate gyrus, a fluorescent microscope (Nikon eclipse 80i, Nikon Instruments, Melville, NY, USA) with a digital camera was used (× 20 objective).
3
FGF2 and DCX Immunostaining in Mouse Brain
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