Immunohistochemical techniques were utilized for assessment of the presence of Bcl-2, Bax, and cleaved caspase-3 [18 ,19 (link)]. Briefly, endogenous peroxidases in the sections were blocked with H2O2, and the sections were then blocked with bovine serum albumin and normal goat serum. Rabbit polyclonal anti-Bcl-2, anti-Bax (1:50 dilution, Santa Cruz Biotechnology Inc., Santa Cruz, CA), or anti-cleaved caspase-3 primary antibody (1:50 dilution, Cell Signaling, Beverly, MA) was added, and the sections were stained using a Vectastain Elite ABC kit (Vector Laboratories, Burlington, ON), in which color was developed with 3,3'-diaminobenzidine tetrahydrochloride (DAB; Roche, Mannheim, Germany). The sections were evaluated at a 200× magnification. The intensity of DAB staining was measured using Image J software. Relative values for the intensity were used by setting the value for the sham group at 1.
3 3 diaminobenzidine tetrahydrochloride dab
Manufactured by Roche
Sourced in Germany
3,3'-diaminobenzidine tetrahydrochloride (DAB) is a chromogenic substrate used in immunohistochemistry and other biochemical assays to detect the presence of target analytes. It produces a brown precipitate upon reaction with horseradish peroxidase (HRP), a commonly used enzyme label in these applications.
Automatically generated - may contain errors
2 protocols using 3 3 diaminobenzidine tetrahydrochloride dab
1
Immunohistochemical Analysis of Apoptosis Markers
2
TUNEL Assay for In Situ Cell Death Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An in situ cell death detection kit, POD (11684817910, Roche, Germany) was used for the TUNEL staining, and the kit protocol was applied following the dewaxing and dehydration. Briefly, dewaxed tissue sections were predigested with proteinase K for 20 min and incubated in PBS containing 3% H2O2 for 10 min to block the endogenous peroxidase activity. After two washes with PBS for 5 min each, the sections were incubated with the TUNEL reaction mixture, fluorescein-dUTP (in situ cell death detection kit, POD, Roche, Germany), for 60 min at 37 °C. The sections were then rinsed three times with PBS for 5 min each and incubated with secondary antifluorescein-POD-conjugate for 30 min. After three washes with PBS for 5 min each, 3, 3′-diaminobenzidine tetrahydrochloride (DAB; 11,718,096,001, Roche, Germany) –H2O2 chromogen was added on the sections and was counterstained with hematoxylin. After rinsing and mounting, the stained images were observed and captured using an inverted light microscope.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!