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Anti neurofilament rabbit polyclonal antibody

Manufactured by Merck Group
Sourced in United States

The Anti-neurofilament rabbit polyclonal antibody is a laboratory reagent used to detect and study neurofilament proteins, which are structural components of neurons. This antibody is generated by immunizing rabbits with neurofilament proteins, and it can be used in various analytical techniques, such as Western blotting and immunohistochemistry, to identify and visualize neurofilaments in biological samples.

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3 protocols using anti neurofilament rabbit polyclonal antibody

1

Immunofluorescent Labeling of Neural Markers

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After washing, cross‐tissue sections were exposed to triton x‐100 (0.3%), and blocked in 10% goat serum for 45 min at room temperature. Tissue sections were then placed overnight in primary antibodies including anti‐glial fibrillary acidic protein (GFAP) monoclonal antibody (1:150, Millipore, Germany), anti‐S100β cell monoclonal antibody (1:400, Cosmo Bio Co., Japan) and anti‐neurofilament rabbit polyclonal antibody (1:400, Sigma, USA). The sections were incubated for 2 h with FITC secondary antibodies at 1:400 dilution. The sections were evaluated using a fluorescing microscope, and the intensity was calculated for each image (Image J software 1.43U). The coefficient of variation was then measured (20% for all groups).
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2

Immunofluorescence Labeling of Neurofilaments

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Immunofluorescence for neurofilament was performed according to the previously described method (Jung et al., 2016 (link)). The sections were washed 3 times with PBS, and then exposed to 0.1% bovine serum albumin solution containing 0.1% Tween 20 in PBS for 4 hr at room temperature. Next, the sections were incubated overnight with a solution containing primary antibodies as antineurofilament rabbit polyclonal antibody (1:400, Sigma Chemical Co., St. Louis, MO, USA) for axons, After washing, the sections were incubated 2 hr with secondary antibodies as rhodamine-goat anti-rabbit secondary antibodies (1:800; Molecular, Probes, Eugene, CA, USA) for neurofilament. The sections were mounted and examined by a fluorescence microscope (Nikon Eclipse 50i, Nikon Inc., Melville, OR, USA).
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3

Immunofluorescence Staining of Neural Markers

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For immunofluorescence, the sections were washed 3 times with PBS, and then exposed to 0.1% bovine serum albumin solution containing 0.1% Tween 20 in PBS for 4 hr at room temperature. Next, sections were incubated overnight with a solution containing primary antibodies as follows: antiglial fibrillary acidic protein monoclonal antibody (1:400, Santa Cruz Biotechnology, Santa Cruz, CA, USA) for astrocyte, anti-neurofilament rabbit polyclonal antibody (1:400, Sigma Chemical Co., St. Louis, MO, USA) for axons, anti-S100β cell monoclonal antibody (1:400, Cosmo Bio Co., Tokyo, Japan) for Schwann cells, and anti-BDNF rabbit polyclonal antibody (1:400, Santa Cruz Biotechnology) for BDNF. After washing, the sections were incubated 2 hr with secondary antibodies as follows: fluorescein isothiocyanate anti-mouse secondary antibody (1:400, Jackson ImmunoResearch Laboratories, West Grove, PA, USA) for neurofilament and Schwann cells or CY3 anti-rabbit secondary antibody (1:800, Jackson ImmunoResearch Laboratories) for astrocyte and BDNF. The sections were mounted and examined by a fluorescence microscope (Nikon Eclipse 50i, Nikon Inc., Melville, NY, USA).
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