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).
Antigen retrieval reagent acidic buffer ph 6.0
Manufactured by R&D Systems
The Antigen Retrieval Reagent-Acidic buffer (pH 6.0) is a laboratory reagent designed to facilitate the unmasking of antigenic sites in tissue samples. It is a buffered solution with a pH of 6.0 that is used in the process of antigen retrieval, a critical step in immunohistochemistry (IHC) and other applications where the detection of specific proteins or molecules is required.
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2 protocols using antigen retrieval reagent acidic buffer ph 6.0
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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