For immunohistochemistry analysis of human skin tissue samples, formalin-fixed paraffin-embedded sections were deparaffinized with CitriSolv Hybrid (Decon Labs) and rehydrated by sequentially incubating in 100%, 95%, 70%, and 50% ethanol in water. To expose antigenic epitopes, heat-induced antigen retrieval was performed using Antigen Retrieval Reagent-Acidic buffer (pH 6.0) (R&D) for 20min at 95°C. After quenching endogenous peroxidase activity by 3% hydrogen peroxide, sections were incubated with the following primary antibodies at 4 °C overnight (see Supplementary Table 8 ): NFIB (1:100), NFIX (1:100), K14 (1:2000). After several washes with PBS, primary antibody staining was visualized using peroxidase-conjugated anti-rabbit IgG (1:1000) followed by the DAB substrate kit for peroxidase visualization of secondary antibodies (Vector Laboratories).
Dab substrate kit for peroxidase visualization of secondary antibodies
Manufactured by Vector Laboratories
The DAB substrate kit is a reagent used for the visualization of peroxidase-labeled secondary antibodies in immunohistochemical and immunocytochemical applications. The kit provides the necessary components to produce a brown, insoluble reaction product at the site of the target antigen.
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2 protocols using dab substrate kit for peroxidase visualization of secondary antibodies
1
Immunohistochemistry Analysis of Human Skin Tissue
2
Immunohistochemistry Analysis of Human Skin Tissue
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For immunohistochemistry analysis of human skin tissue samples, formalin-fixed paraffin-embedded sections were deparaffinized with CitriSolv Hybrid (Decon Labs) and rehydrated by sequentially incubating in 100%, 95%, 70%, and 50% ethanol in water. To expose antigenic epitopes, heat-induced antigen retrieval was performed using Antigen Retrieval Reagent-Acidic buffer (pH 6.0) (R&D) for 20min at 95°C. After quenching endogenous peroxidase activity by 3% hydrogen peroxide, sections were incubated with the following primary antibodies at 4 °C overnight (see Supplementary Table 8 ): NFIB (1:100), NFIX (1:100), K14 (1:2000). After several washes with PBS, primary antibody staining was visualized using peroxidase-conjugated anti-rabbit IgG (1:1000) followed by the DAB substrate kit for peroxidase visualization of secondary antibodies (Vector Laboratories).
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