Immunohistochemistry for VEGF was performed by incubating three-micrometer paraffin sections with normal donkey serum for 1 h (D9663, Sigma-Aldrich), and then goat anti-rat VEGF (1:500, 564-RV, R&D Systems, Minneapolis, Minnesota, USA) overnight at 4 °C. A negative control without the primary antibody and an isotype IgG control were included. Sections were washed with PBS, incubated for 30 min with biotin-conjugated donkey anti-goat IgG (1:500, 705-065-147, Jackson ImmunoResearch, Pennsylvania, USA), washed with PBS, and then incubated with the Vectastain ABC standard kit (Vector Laboratories, Pennsylvania, USA) for 30 to 45 min and liquid DAB+substrate chromagen system (Dakocytomation) for 15 s. The sections were counterstained with Mayer’s Haematoxylin and coverslipped. Quantitation was performed as described for GFAP. Data are presented as the percentage of VEGF immunolabeling per field of retina. Four to five rats per group were evaluated.
Dab substrate chromagen system
Manufactured by Agilent Technologies
Sourced in United States
The DAB+substrate chromagen system is a laboratory equipment product that provides a chromogenic detection method for immunohistochemistry and in situ hybridization applications. It generates a brown color reaction when exposed to the target analyte, enabling visualization and analysis.
Automatically generated - may contain errors
2 protocols using dab substrate chromagen system
1
Quantifying Retinal VEGF Expression
2
Autopsy Findings in COVID-19 Patients
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Autopsies of 23 patients infected with Sars-CoV-2, documented by PCR on a pre or postmortem nasopharyngeal swab, were examined. Autopsies were consented for and performed on the clinical service with complete examination of chest organs and in-situ sampling of remaining organs and tissues, with histology performed on all sites. Lung paraffin sections from COVID-19 autopsy patients were either stained with Hematoxylin and Eosin, or processed as follows. After deparaffinization and rehydration, the sections were immersed in antigen retrieval solution (DAKO) for 30 min at 98°C. For IgM staining, the sections were blocked with goat serum (30 minutes at room temperature), followed by incubation with horseradish peroxidase labeled goat anti-human IgM (Jackson ImmunoResearch, cat # 109-035-043) diluted 1:500. Visualization was performed with a liquid DAB substrate-chromagen system (DAKO) and the sections were counterstained with hematoxylin before mounting.
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