Histological sections of the rats given an i.v. injection of aPS-PT mAb, in which thrombi were present, were subjected to immunohistochemistry for rat IgM. For this purpose, we used horseradish peroxidase-conjugated goat anti-rat IgM antibodies (Abcam, Cambridge, UK) and the diaminobenzidine substrate kit (Sigma-Aldrich). Sections of the rats given an i.v. injection of calf thymus-derived histones only were stained simultaneously as controls.
Diaminobenzidine substrate kit
Manufactured by Merck Group
Sourced in United Kingdom, United States
The Diaminobenzidine substrate kit is a laboratory reagent used in immunohistochemistry and enzyme-linked immunosorbent assays (ELISA) to detect the presence of specific proteins or antigens in biological samples. The kit contains a stable diaminobenzidine solution that produces a brown color upon reaction with an enzyme-labeled detection system. This colorimetric reaction allows for the visualization and localization of the target analyte within the sample.
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2 protocols using diaminobenzidine substrate kit
1
Immunohistochemical Analysis of Rat IgM
2
Immunofluorescence and Immunohistochemistry Protocols
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The transfected cells grown on the cover slides were washed with cold PBS, fixed with 4% paraformaldehyde, and permeabilized with 0.2% Triton-X. After four washing with PBST, cells were further stained by DAPI (Thermo Fisher Scientific, U.S.) and imaged with a Zeiss LSM710 microscope.
For IHC analysis, paraffin-embedded tissues were fixed on slides and then blocked with 5% donkey serum (Gibco, U.S.) for 45 min, followed by incubation with anti-pNF90-Ser382 antibody (Youke, Shanghai, China) or anti-Ki67 antibody (Abcam, U.K.) overnight at 4 °C. The horseradish peroxidase-conjugated secondary antibody was used and the Diaminobenzidine Substrate Kit (Sigma, U.S.) was applied for signal detection. All tissue sections were counterstained with hematoxylin (Sigma, U.S.).
For IHC analysis, paraffin-embedded tissues were fixed on slides and then blocked with 5% donkey serum (Gibco, U.S.) for 45 min, followed by incubation with anti-pNF90-Ser382 antibody (Youke, Shanghai, China) or anti-Ki67 antibody (Abcam, U.K.) overnight at 4 °C. The horseradish peroxidase-conjugated secondary antibody was used and the Diaminobenzidine Substrate Kit (Sigma, U.S.) was applied for signal detection. All tissue sections were counterstained with hematoxylin (Sigma, U.S.).
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