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Anti bdnf rabbit polyclonal antibody

Manufactured by Santa Cruz Biotechnology
Sourced in United States

The Anti-BDNF rabbit polyclonal antibody is a laboratory reagent designed to detect the presence of brain-derived neurotrophic factor (BDNF) in biological samples. It is a protein-based tool used in various research applications, such as Western blotting, immunohistochemistry, and ELISA, to study the expression and distribution of BDNF in different tissues or cell types.

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4 protocols using anti bdnf rabbit polyclonal antibody

1

Western Blot Analysis of BDNF in Spinal Cord

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Protein extracts from spinal cord tissue were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Protein separation was performed using a 10% polyacrylamide with 0.05% bis-acrylamide. Proteins were then transferred to nitrocellulose and the blots were probed with anti-BDNF rabbit polyclonal antibody (1:1,000, Santa Cruz Biotechnology) and anti-β-actin mouse monoclonal antibody (1:3,000, Santa Cruz Biotechnology). Peroxidase anti-rabbit IgG (1:5,000, Vector Laboratories, Burlingame, CA, USA), and peroxidase anti-mouse IgG (1:5,000, Vector Laboratories) were used as a secondary antibodies. Immunoreactivity was detected by enhanced chemiluminescence (Santa Cruz Biotechnology). Film autoradiograms were exposed from 5 to 15 min.
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2

Western Blot Analysis of BDNF and TrkB

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Western blot analysis for BDNF and tyrosine kinase B (TrkB) was performed, according to a previously described method (Kim et al., 2010 (link)). Proteins were then transferred to nitrocellulose and the blots were probed with anti-BDNF rabbit polyclonal antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or anti-TrkB rabbit polyclonal antibody (1:1,000, Santa Cruz Biotechnology). Peroxidase anti-rabbit IgG (1:5,000, Vector Laboratories) was used as the secondary antibodies. Immunoreactivity was detected by enhanced chemiluminescence detection kit (Santa Cruz Biotechnology).
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3

Quantifying BDNF and Synapsin-I in Spinal Cord

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Western blot for BDNF and synapsin-I was performed according to the previously described method (Jung et al., 2016 (link)). The spinal cord tissues covering approximately 1 cm of the rostral and caudal spinal cord at the injury area were prepared and washed with ice-cold PBS and sonicated in 400–600 mL of Triton lysis buffer. Protein separation was performed using a 10% polyacrylamide with 0.05% bis-acrylamide. Proteins were then transferred to nitrocellulose and the blots were probed with anti-BDNF rabbit polyclonal antibody (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-synapsin-I rabbit monoclonal antibody (1:1,000, Cell Signaling Technology, Beverly, MA, USA), and anti-β-actin mouse monoclonal antibody (1:3,000, Santa Cruz Biotechnology). Peroxidase anti-rabbit IgG (1:5,000; 1:5,000 Vector Laboratories, Burlingame, CA, USA), and peroxidase anti-mouse IgG (1:5,000, Vector Laboratories) were used as the secondary antibodies. Immunoreactivity was detected by enhanced chemiluminescence (Santa Cruz Biotechnology).
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4

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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