The tissue expressions of IL-17 and S100A7 were assessed with an immunohistochemical method. The immunohistochemical reactions were performed on 4 µmformalin-fixed and paraffin-embedded (FFPE) biopsy specimens according to the manufacturer’s protocol, using mouse anti-human IL-17 polyclonal antibodies (catalogue number PA1-84183; Invitrogen, USA), mouse anti-human psoriasin (S100A7) monoclonal antibodies (catalogue number ab13680; Abcam, UK), with positive and negative control stainings. Detection was performed on the Dako platform using the Dako RDS AP kit (Dako Real Detection System Alkaline Phosphatase Real Rabbit/Mouse, catalogue number K5005). The preparations were assessed using Zeiss Axio Imager A2 light microscope with video track and Zeiss AxioVision software. The expression was considered as positive when membrane or cell cytoplasmic immunoreactivity was observed. Positively stained cells were counted in 10 fields of view for each specimen preparation at 200x magnification, and the mean value for each preparation was calculated.
Dako rds ap kit
Manufactured by Agilent Technologies
The Dako RDS AP kit is a reagent kit used in immunohistochemistry (IHC) procedures. It contains the necessary components to perform a labeled streptavidin-biotin (LSAB) detection system. The kit is designed to detect primary antibodies bound to target antigens in tissue sections or cell preparations.
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2 protocols using dako rds ap kit
1
Immunohistochemical Analysis of IL-17 and S100A7
2
Quantification of IL-17 Expression
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The immunohistochemical assay was performed using mouse anti-human IL-17 polyclonal antibodies (dilution 0.25 μg; catalogue number PA1-84183; Invitrogen, USA). All determinations were made in accordance with the manufacturer's protocol, with positive and negative control stainings. Visualization was carried out using the Dako RDS AP kit (Dako Real Detection System Alkaine Phosphotase Real Rabbit/Mouse, catalogue number K5005). The preparations were evaluated in a Zeiss Axio Imager A2 light microscope using a video track and Zeiss AxioVision software. The expression was considered to be positive when cell cytoplasmic or membrane immunoreactivity was observed. Positively stained cells were counted in 10 fields of view for each preparation at 200x magnification, and the mean value for each preparation was calculated.
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