Microwave Heating and Ultrasound Antigen Retrieval Method

COMBINED MICROWAVE HEATING AND ULTRASOUND ANTIGEN RETRIEVAL METHOD 

The combined heat-induced epitope retrieval (HIER) and sonication-induced epitope retrieval (SIER) methods have been employed for retrieving cyclin Dl/bc 1-1 epitope in 

The slides are placed in a plastic microwavable rack and immersed in a microwavable staining dish filled with 200 ml of 0.01 M citrate buffer (pH 6.0). Blank slides are added to the rack to maintain a uniform volume in the container. The container is covered at an angle with the lid and heated twice for 5 min per cycle at 800 W in a microwave oven. Distilled water (50ml) is added after the first heating cycle to cool at room at room temperature for 10 min. 

The buffer (-800 ml) is heated to boiling in the microwave oven and poured into an 80-W ultrasonic cleaner (Bransonic 12, Branson Cleaning Equipment Co., Shelton, CT). The slide rack is sonicated for 1 min and then cooled in buffer in the staining container for an additional 10 min. 

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