Immunofluorescence for brain slices was performed on fixed frozen sections as previously described (Hu et al., 2009 (link)). Twenty-four hours after SAH, rats were deeply anesthetized and transcardially perfused with phosphate-buffered solution (PBS) and 10% formalin. Rats’ brains were rapidly isolated and postfixed in 10% formalin for 24 hours and then in 30% sucrose for 3 days. Coronal brain sections (10 μm) were obtained with the help of cryostat (Leica CM3050S-3-1-1, Bannockburn, IL) and permeabilized with 0.3% Triton X-100 in PBS for 30 minutes. Sections were blocked with 5% donkey serum for 1 hour and incubated at 4 °C overnight with primary antibodies: anti-phosphorylated TrkB (Abcam, Cambridge, MA) and anti-neuronal nuclei (NeuN) (Millipore, Temecula, CA), followed by fluorescein isothiocyanate (FITC)- and Texas Red-conjugated secondary antibodies (Jackson Immunoresearch, West Grove, PA) for 2 hours at room temperature. The colocalization of p-TrkB with the marker of neuron was examined by fluorescent microscope (Olympus OX51, Tokyo, Japan).
Anti neuronal nuclei neun
Manufactured by Merck Group
Anti-neuronal nuclei (NeuN) is a monoclonal antibody that recognizes a neuronal nuclear protein expressed in most neuronal cell types. It is commonly used as a marker for identifying and quantifying mature neurons in the central and peripheral nervous systems.
Automatically generated - may contain errors
Lab products found in correlation
Anti phosphorylatedtrkb, by Abcam (1 mentions)
Cm3050s 3 1 1, by Leica (1 mentions)
Tissue culture reagents, by Thermo Fisher Scientific (1 mentions)
Anti bace1, by Cell Signaling Technology (1 mentions)
6e10 against aβ1 16, by Fortrea (1 mentions)
Anti β actin, by Abcam (1 mentions)
Anti psd95, by Cell Signaling Technology (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
3 protocols using anti neuronal nuclei neun
1
Immunofluorescence Analysis of Phosphorylated TrkB in Brain Slices
2
Protein Detection Immunohistochemistry Protocol
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The antibodies used for detection of proteins of interest were 6E10 (against Aβ1-16) from Covance, R1(57) against the carboxy terminus of APP was a kind gift from Dr P. Mehta (NYS Institute for Basic Research in Developmental Disabilities); anti-BACE1 was from Cell Signaling; anti-apolipoprotein E (ApoE) and anti-neprilysin CD10 from Santa Cruz; anti-ionized calcium-binding adapter molecule 1 (IBA1) from Wako; anti-glial fibrillary acidic protein (GFAP) (clone 2.2B10), anti-Aβ (6C3), and anti-neuronal nuclei (NeuN) were from Millipore; and anti-insulin degrading enzyme (IDE), Aldehyde dehydrogenase 1A (Aldh1a1), and anti-β-actin were from Abcam. Tissue culture reagents were purchased from Invitrogen and Millipore, and all other reagents were purchased from Sigma, unless stated otherwise.
3
Western Blot Analysis of MAM-Induced Neurochemical Changes
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For western blot analysis, bilateral cortical tissues from bregma to posterior hippocampal areas without hippocampus (anterior posterior 0 to -5 mm) were obtained from control rats (n = 12), MAM-exposed rats (n = 12), rhIGF-1-treated MAM-exposed rats (n = 11) and VEH-treated MAM-exposed rats (n = 9) at P15. The following primary antibodies were used: anti-AMPA receptor 1 (AMPAR1), AMPAR2, AMPAR3, and AMPAR4 (Cell signaling, Technology, Inc.); anti-calcium/calmodulin-dependent protein kinase II (CaMKII) (Cell signaling, Technology, Inc.); anti-glutamic acid decarboxylase 67 (GAD67) and GAD65 (Millipore, Technology, Inc.); anti-neuronal nuclei (NeuN; Millipore, Technology, Inc.); anti-N-methyl-D-aspartate receptor 1 (NMDAR1), NMDAR2A, and NMDAR2B (Cell signaling, Technology, Inc.); anti-PSD95 (Cell signaling, Technology, Inc.); Anti-β-actin (Santa Cruz Biotechnology, Inc.).
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