EBER (Epstein-Barr virus-encoded small RNAs) ISH was performed using a Ventana Benchmark ISH system (Ventana ISH iView kit, Ventana Corporation, Tucson, AZ, USA). Paraffin-embedded tissue sections were deparaffinized using EZ Prep buffer (Ventana Corporation), and then digested with Protease I for 4 min. Probes were added to the sample, and then denaturation was performed at 85°C (10 min), followed by hybridization at 37°C (1 hour). The probes labeled with fluorescein contained a cocktail of oligonucleotides dissolved in a formamide-based diluent. After hybridization, tissues were washed 3 times using 2x saline sodium citrate buffer at 57°C. Incubation with anti-fluorescein monoclonal antibody was performed for 20 min and then the alkaline blue detection kit (Ventana Corporation) was used according to the manufacturer's protocol. The slides were counterstained with nuclear fast red (Ventana Corporation) for 10 min.
Alkaline blue detection kit
Manufactured by Roche
The Alkaline Blue detection kit is a laboratory equipment product designed to detect the presence of alkaline phosphatase in various samples. The kit provides the necessary reagents and components to perform this specific analysis.
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Lab products found in correlation
3 protocols using alkaline blue detection kit
1
EBER ISH Detection in Tissue Sections
2
EBER ISH Protocol for Tissue Analysis
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EBER ISH was performed with a Ventana BenchMark in situ hybridization system (ISH iView kit, Ventana, Tucson, AZ, USA). Paraffin-embedded tissue sections were deparaffinzed with EZ Prep buffer (Ventana), and then digested with protease I for 4 min. Probes were applied and then denaturation was performed at 85 °C (10 min), followed by hybridization at 37 °C (1 h). The probes labeled with fluorescein contained a cocktail of oligonucleotides dissolved in a formamide-based diluent. After hybridization, tissues were washed 3 times with 29 saline sodium citrate buffer at 57 °C. Incubation with antifluorescein monoclonal antibody was performed for 20 min and then an Alkaline Blue detection kit (Ventana) was used according to the manufacturer's protocol. The slides were counterstained with Nuclear Fast Red for 10 min.
3
EBER ISH for EBV Detection
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Epstein-Barr virus-encoded RNA in situ hybridization (EBER ISH) was performed with a Ventana Bench Mark system (ISH iView kit; Ventana, Tucson, AZ, USA). Paraffin-embedded tissue sections were deparaffinized with EZ Prep buffer (Ventana) and digested with protease I for 4 min. Then, probes for EBER were denatured at 85 °C for 10 min and then hybridized at 37 °C for 1 h. After hybridization, the tissues were washed with 29 SSC buffer at 57 °C. Incubation with anti-fluorescein monoclonal antibodies was then performed for 20 min, and then an Alkaline Blue detection kit (Ventana) was used according to the manufacturer's protocol. The slides were counterstained with Nuclear Fast Red for 10 min.
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