In the same manner, as described below, immunohistochemistry for DCX was performed (Park et al., 2019 (link)). The sections were treated with PBS for 10 min and then treated again with 1% H2O2 for 20 min. The sections were treated with goat anti-DCX antibody (1:1,000; Oncogene Research Product, Cambridge, UK) for 2 hr. The sections were treated with the biotinylated goat secondary antibody (1:500; Vector Laboratories, Burlingame, CA, USA) for another 1 hr, washed, and treated with ABC complex (1:100; Vector Laboratories). Using the 0.03% diaminobenzidine, labeling was visualized and the sections were mounted onto gelatin-coated slides. After air drying the slides at room temperature overnight, the coverslips were mounted using Permount (Thermo Fisher Scientific Inc., Waltham, MA, USA).
Biotinylated goat secondary antibody
Manufactured by Vector Laboratories
Sourced in United States
Biotinylated goat secondary antibody is a reagent used in various immunoassay techniques. It is a goat-derived antibody that has been chemically modified to include biotin, a small molecule that can bind to the protein streptavidin. This allows the antibody to be detected or manipulated in experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti dcx antibody, by Santa Cruz Biotechnology (4 mentions)
Vector elite abc kit, by Vector Laboratories (4 mentions)
Permount, by Thermo Fisher Scientific (4 mentions)
Avidin peroxidase complex, by Vector Laboratories (2 mentions)
Abc complex, by Vector Laboratories (1 mentions)
Ab19027, by Abcam (1 mentions)
Immpact dab, by Vector Laboratories (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine dab substrate kit, by Vector Laboratories (1 mentions)
Normal goat serum (ngs), by Vector Laboratories (1 mentions)
Prolong gold with dapi, by Thermo Fisher Scientific (1 mentions)
Normal goat serum ngs, by Vector Laboratories (1 mentions)
9 protocols using biotinylated goat secondary antibody
1
DCX Immunohistochemistry Protocol
2
DCX Immunohistochemistry Protocol
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According to a previously described method [13 (link)], DCX immunohistochemistry was conducted. The sections were incubated by goat anti-DCX antibody (1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight and next incubated by biotinylated goat secondary antibody (1:200; Vector Laboratories) for another 1 hour. Then, the bound secondary antibody was amplified with a Vector Elite ABC kit (1:200; Vector Laboratories). The 0.02% diaminobenzidine was used to visualize the antibody-biotin-avidin-peroxidase complex. After dehydration through a gradient of ethanol, the slides were mounted with coverslips using PermountTM mounting media (Thermo Fisher Scientific).
3
Immunohistochemistry for Doublecortin (DCX)
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Immunohistochemistry for DCX was performed, according to the previous method (Park and Kim, 2017 (link)). The sections were incubated in PBS for 10 min, washed three times in PBS, and then incubated in 1% hydrogen peroxide for 20 min. The sections were incubated 2 hr with goat anti-DCX antibody (1:1,000; Oncogene Research Product, Cambridge, UK). The sections were then incubated with the biotinylated goat secondary antibody (1:500; Vector Laboratories, Burlingame, CA, USA) for another 1 hr, washed, and incubated in ABC complex (Vector Elite ABC kit; 1:100; Vector Laboratories). Labeling was visualized using 0.03% diaminobenzidine, and the sections were mounted onto gelatin-coated slides. The slides were air-dried overnight at room temperature, and the coverslips were mounted using Permount (Thermo Fisher Scientific Inc., Waltham, MA, USA).
4
Immunohistochemical Staining of DCX
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Immunohistochemistry for DCX was performed as previously described method (Park et al., 2019 (link)). The sections were incubated in PBS for 10 min, washed three times in PBS, and then incubated in 1% hydrogen peroxide for 20 min. The sections were incubated 2 hr with goat anti-DCX antibody (1:1,000; Oncogene Research Product). The sections were then incubated with the biotinylated goat secondary antibody (1:500; Vector Laboratories) for another 1 hr, washed, and incubated in ABC complex (Vector Elite ABC kit; 1:100; Vector Laboratories). Labeling was visualized using 0.03% DAB, and the sections were mounted onto gelatin-coated slides. The slides were air-dried overnight at room temperature, and the coverslips were mounted using Permount (Thermo Fisher Scientific Inc.)
5
Immunohistochemical Detection of Cathepsin K
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Tissue sections were deparaffinized and rehydrated, then endogenous peroxidase was blocked with 3% H2O2/PBS solution. After blocking with 2% goat serum, sections were incubated with an antibody against Cathepsin K (ab19027, Abcam) overnight at 4 °C. After washing with PBS, sections were incubated with biotinylated goat secondary antibody (Vector Laboratories, Burlingame, CA, USA) for 60 min at room temperature followed by treatment with Vectastain elite ABC kit (Vector Laboratories). Sections were colored using ImmPACT DAB (Vector Laboratories) and counterstained with hematoxylin.
6
Immunohistochemical Detection of DCX
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DCX immunohistochemistry was assessed using a previously described method (Shin et al., 2013 (link)). The sections were treated with 3% H2O2 for 30 min at room temperature. Next, the sections were blocked with 10% normal rabbit serum in PBS with 1% bovine serum albumin and 0.2% Triton X-100 for 1 hr and incubated with goat anti-DCX antibody (1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight at 4°C. After washing 3 times, the sections were incubated with biotinylated goat secondary antibody (1:200; Vector Laboratories) for 1 hr, followed by incubation for 1 hr with avidin-peroxidase complex (1:100; Vector Laboratories). For immunostaining, the sections were visualized in 50-mM Tris–HCl (pH, 7.6) containing 0.03% DAB and 0.03% H2O2. Then, the tissue samples were rinsed with PBS three times and mounted on gelatin-coated slides. The slides were dehydrated through alcohol and coverslipped with Permount (Fisher Scientific Inc.).
7
Visualizing Hippocampal Cell Differentiation
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In order to visualize cell differentiation, immunohistochemistry was performed to detect dentate gyrus (DG), and doublecortin (DCX) in the DG region of the hippocampus. The sections were incubated in PBS for 10 min and then washed three times for 3 min in PBS. The sections were then incubated in 1% H2O2 for 15 to 30 min. Sections were obtained from each brain and incubated overnight with goat anti-DCX antibody (1:500; Santa Cruz Biotechnology, Dallas, TX, USA) followed by incubation with biotinylated goat secondary antibody (1:200; Vector Laboratories, Burlingame, VT, USA) for another 90 min and then washed and incubated in Vector Elite ABC Kit (1:100; Vector Laboratories). Antibody-biotin–avidin–peroxidase complexes were visualized using the 3,3, Diaminobenzidine (DAB) Substrate Kit (Vector Laboratories). The slides were air-dried overnight at room temperature, and the coverslips were mounted using Permount.
8
Doublecortin Immunohistochemistry Protocol
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Following a method described elsewhere [18 (link)], doublecortin (DCX) immunohistochemistry was performed. After the sections were incubated in 3% H2O2 for 30 minutes at room temperature, they were blocked by 10% normal rabbit serum in PBS with 0.2% Triton X-100 and 1% bovine serum albumin for 1 hour. The section was incubated in goat anti-DCX antibody (1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4℃ overnight. After washing 3 times, the sections were treated with a biotinylated goat secondary antibody (1:200; Vector Laboratories) for 1 hour, followed by another hour with avidin–peroxidase complex (1:100; Vector Laboratories). The sections were visualized in 50mM Tris-HCl (pH, 7.6) containing 0.03% hydrogen peroxide and 0.03% DAB for immunostaining. The slides were dehydrated by alcohol and mounted using Permount coverslips (Thermo Fisher Scientific Inc.).
9
Quantifying DNA Methylation Variants
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