Sigmafast dab tablet

The following antibodies were used for immunohistochemistry anti-Ki67 (DAKO) and anti-YAP (CST). Unstained 5μm paraffin sections were dewaxed in Safeclear II (Fisher Scientific, PA), hydrated through graded alcohols and distilled water, and washed three times with PBS. Antigens were retrieved using or 10mM citrate buffer boiled in a microwave for 20 min (2 min at 100% power and 18 min at 10% power). The slides were allowed to cool down for 30 min at room temperature, rinsed twice with PBS, incubated in 3% hydrogen peroxide in PBS for 10 min to quench the endogenous peroxidase. The sections were then sequentially washed in distilled water and PBS, incubated in blocking solution (2.5% bovine serum albumin in PBS) for 30 min at room temperature. Excess solution was discarded and the primary antibodies were applied diluted in blocking solution at 4°C overnight. After washing with PBS, the slides were sequentially incubated with the biotinylated secondary antibody (1:400) (Vector Laboratories, CA) for 30 min and with the avidin-biotin complex, reconstituted according to the instruction of the manufacturer in PBS (Vector Stain Elite, ABC kit) (Vector Laboratories, CA), for 30 min at room temperature. The slides were developed in 3,3-diaminobenzidine (Sigma FASTDAB tablet) (Sigma Chemical, MO) diluted in distilled water under a microscope.