SP immunohistochemistry assay for different receptor subunits was performed. Four to six slides from each rat were deparaffinated with xylene and alcohol of different concentration and incubated with 3% H2O2. Antigens of sections were retrieved with 0.125% trypsin solution. Sections were rinsed in Tris-HCl-buffered saline and preincubated with normal goat serum (SP-9001, ZSGB-BIO, China), followed by incubation with the primary antibodies, anti-GAD65 + GAD67 (Abcam), anti-GAT1 (Abcam), anti-BDNF (Abcam), and anti-TrkB (Abcam), at 4°C overnight. Subsequently, they were incubated with horseradish peroxidase-coupled goat anti-rabbit secondary antibodies (SP-9001, ZSGB-BIO, China) for 60 min at 37°C temperature. Finally, the sections were reacted with 0.4 mM 3,30-diaminobenzidine (ZSGB-BIO, China) and 0.01% H2O2 for 10–15 min. After each incubation step, except the preincubation, three 5 min washes with Tris-HCl-buffered saline were performed.
3 30 diaminobenzidine
Manufactured by ZSGB-BIO
Sourced in China
3,30-diaminobenzidine is a chromogenic substrate used in immunohistochemistry and histochemistry applications. It produces a brown precipitate upon oxidation, allowing for the visualization of target proteins or enzymes in biological samples.
Automatically generated - may contain errors
3 protocols using 3 30 diaminobenzidine
1
Immunohistochemical Analysis of Neurotransmitter Receptors
2
Immunohistochemical Staining of Lung Tissue
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After deparaffinization and hydration, a citrate solution was used for antigen retrieval of the lung tissue slides. After blocking with 0.5% BSA, the slides were incubated with primary antibodies (dilution at 1:100) at 4 ℃ overnight. Then they were incubated with a Rabbit Two-step Detection Kit (PV-9001, ZSGB-BIO, P.R.China), and a chromogenic reaction with 3,30-diaminobenzidine (ZLI-9018, ZSGB-BIO, P.R.China) was performed. Then images were captured under a light microscope (Nikon, Japan).
3
Immunohistochemical Analysis of mTOR and Ki67
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Tissue sections were deparaffinized in xylene and rehydrated in descending concentrations of ethanol, followed by antigen retrieval in sodium citrate buffer for 10 min in a microwave oven at 100°C and cooling to room temperature. The experiments were carried out using Histostain-Plus Kits (ZSGB-BIO, Beijing, China), and a chromogenic reaction was carried out with 3,30-diaminobenzidine (ZSGB-BIO, Beijing, China) according to the manufacturer's protocol. The primary antibodies used were for mTOR (#2983; Cell Signal Technology, Boston, MA, USA) and Ki67 (ab16667; Abcam, Cambridge, US), both at a 1:200 dilution (Proteintech, Chicago, IL, USA). The sections were counterstained with haematoxylin and mounted with resinene. The images were collected on an Olympus microscope (BX40, Tokyo, Japan).
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