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Biotinylated goat anti rabbit igg

Manufactured by Zymo Research
Sourced in United States

Biotinylated goat anti-rabbit IgG is a secondary antibody that binds to rabbit primary antibodies. It is labeled with biotin, which can be used for various detection and signal amplification applications.

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4 protocols using biotinylated goat anti rabbit igg

1

Immunohistochemical Localization of Fos

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Frozen coronal sections were cut 30 mm thick. After being rinsed in 0.01 M of PBS, they were incubated with blocking buffer (5% goat serum and 0.3% Triton X-100 in PBS) for 30 min and then with rabbit anti-Fos antibody (sc-52, Santa Cruz, CA, USA) at a dilution of 1:2000 for 24 h at 44 °C. Subsequently, the sections were incubated with the biotinylated goat anti-rabbit IgG (Zymed Laboratories, San Francisco, CA, USA) for 1 h at room temperature. The specificities of the staining were tested on the sections in the control rats by omitting the primary antibodies.
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2

Immunohistochemical Analysis of VLPAG

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Rats were perfused with 0.01 M phosphate-buffered saline (PBS, pH 7.4) followed by 250 mL freshly prepared 4% paraformaldehyde in 0.1 M phosphate buffer (4°C). The brains were removed immediately, postfixed for 4 hours, and dehydrated overnight in 20% sucrose in 0.1 M phosphate buffer saline (PBS) at 4°C. Thirty-micrometer successive coronal sections containing the VLPAG were cut and then collected into 0.01 M PBS. Endogenous active enzymes were blocked with 2% goat serum in 0.01 M PBS containing 0.3% Triton X-100 for 1 hour at room temperature. The GFAP-immunoreactive (GFAP-IR), c-Fos-immunoreactive (c-Fos-IR), and p-ERK1/2-immunoreactive (p-ERK 1/2-IR) sections were incubated overnight at 4°C with rabbit anti-GFAP 1:300 (Bioss Antibodies, Beijing, China), rabbit anti-c-Fos (1:500; Santa Cruz Biotechnology, Dallas, TX, USA), and rabbit anti-p-ERK1/2 (1:1000; Cell Signaling Technology, Danvers, MA, USA). Subsequently, the sections were incubated with biotinylated goat anti-rabbit IgG (Zymed Laboratories, San Francisco, CA, USA) for 1 hour at room temperature. The specificities of the staining were tested on the sections in the control rats by omitting the primary antibodies.
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3

Immunohistochemical Staining Protocol

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After several rinses in KPBS, the free floating sections were placed in biotinylated goat anti-rabbit IgG (Zymed, San Francisco, CA) at a dilution of 1:600 in 0.4% Triton X-100 solution for 4 hrs at room temperature. The tissues were rinsed again in KPBS before being placed in an avidin-biotinylated peroxidase complex solution (ABC kit: Vector Laboratories, Burlingame, CA) overnight at 4°C. Following several rinses in KPBS, the sections were reacted with 0.03% diaminobenzidine, 0.008% nickel ammonium sulfate, and 0.0075% hydrogen peroxide in sodium phosphate buffer. The reacted sections were mounted on chrome-alum subbed slides, dehydrated, and coverslipped. The alternate sections were mounted, stained with 0.1% thionin, dehydrated and coverslipped.
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4

Immunohistochemical Analysis of α 1A -AR in Ovary

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For immunohistochemistry (IHC), the ovary sections were dewaxed, rehydrated and treated with 10% normal goat serum in PBS to block nonspecific binding sites. For α 1A -AR staining, an antibody against the adrenergic receptor (1:50; sc-1477, Santa Cruz Biotechnology) was incubated at 4°C for 12 h. In the negative control group, the antibody against the adrenergic receptor was replaced by PBS. The sections were then incubated with a biotinylated goat anti-rabbit IgG (1:200; Zymed Laboratories) and horseradish peroxidase (HRP)-conjugated streptavidin (1:500, Jackson ImmunoResearch Laboratories) at RT for 2 h. Visualization was done using diaminobenzidine (DAB) with hematoxylin counterstaining.
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