The levels of apoptosis were detected using an In Situ Cell Death Detection kit (Roche Diagnostics GmbH, Mannheim, Germany), according to the manufacturer's instructions. Paraffin sections were inactivated using H2O2 solution, prior to incubation with 50 µl TUNEL reaction solution (Roche Diagnostics GmbH) at 37°C for 60 min. Following washing with PBS, the sections were incubated with 50 µl converter-POD working solution (Roche Diagnostics GmbH) at 37°C for 30 min. Following development with 3′-diaminobenzidine (Beijing Solarbio Science & Technology Co., Ltd.), the nuclei were stained with hematoxylin. The sections were observed under a microscope and the images were captured.
3 diaminobenzidine
Manufactured by Solarbio
Sourced in China
3′-diaminobenzidine is a chromogenic substrate used in immunohistochemistry and other bioanalytical techniques. It undergoes a colorimetric reaction when exposed to peroxidase enzymes, resulting in the formation of a brown precipitate that can be visually detected.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using 3 diaminobenzidine
1
TUNEL Assay for Apoptosis Detection
2
Immunohistochemical Analysis of MAP2 Expressing Neurons
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Brain sections were rinsed with PBS, treated with 0.3% H2O2 for 15 min, and rinsed in PBS (three times for 5 min each). Sections were then blocked with 1% goat serum albumin (SL039; Solarbio, Beijing, China) for 10 min at room temperature and incubated overnight at 4 °C with a MAP2 primary antibody (17490-1-AP, 1:100; Proteintech, Rosemont, IL, USA). Sections were rinsed and incubated in the appropriate secondary antibody for 1 h at room temperature. A solution containing 3′-diaminobenzidine (Solarbio, Beijing, China) was added after incubation with the secondary antibodies. Sections were then dehydrated using an alcohol gradient, cleared in xylene, and mounted for microscopic examination (DMI5000M; Leica) to count the number of positive cells.
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