As formerly described by Gendy et al., (2022) [85 (link)], 5 µm slices were sliced into positive charged slides, rehydrated, and heat-retrieved before being incubated with primary anti-TNF- and VEGF for the duration of the night (at a dilution of 1:100). Following washing, tissue sections were incubated for 30 min at room temperature with a 1:1000 dilution of an HRP-labeled secondary antibody before being blocked for endogenous peroxidases. The color was created using a DAB-Substrate Kit. Slides with negative controls were produced by skipping the primary antibody stage. Using an Olympus BX43 microscope, slides were inspected, and an Olympus DP-27 camera was used to take pictures (Tokyo, Japan). Using Cell Sens Dimensions, positive immune staining was measured as an area percentage (Olympus software).
Antibody Cell Dilution MicroscopePeroxidases Sens Tissue Vegf
Corresponding Organization : October 6 University
Other organizations :
Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement, Université de Bordeaux, Unité de Recherche Œnologie, Institut Polytechnique de Bordeaux, National Research Centre, MetaboHUB, Cairo University, Alfaisal University, Al-Azhar University, Menoufia University, Zagazig University, University of Jordan
Incubation time for primary antibodies (overnight)
Incubation time for secondary antibody (30 min)
Room temperature
Olympus BX43 microscope and DP-27 camera (Tokyo, Japan)
Cell Sens Dimensions software (Olympus)
controls
Negative controls: Slides produced by skipping the primary antibody stage
Annotations
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