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t. uc od yo ur pr Total p53 ELISA Kit User’s Manual For Research Use Only, Not for use in diagnostic procedures ith Summary of Procedure Incubate O/N at 37°C in CO2 incubator ca Incubate appropriate time at 37°C in CO2 incubator m Add appropriate amount of the drug for induction of p53 accumulation ew Culture cells in culture flask or dish at 50-70 % confluency th at Harvest the cells by scraping and centrifugation Add 100 µL of diluted cell lysate to the wells rm Incubate for 1 hour at room temp. an u al Make cell lysate by adding extraction buffer and centrifugation se Wash the wells eu Add 100 µL of Primary Antibody Solution (anti-p53 monoclonal antibody) th Incubate for 1hour at room temp. Wash the wells rt o Add 100 µL of Secondary Antibody Solution (HRP conjugated anti-mouse IgG antibody) fe Incubate for 1hour at room temp. re Wash the wells ea se Add 100 µL of Substrate Reagent nl y! Pl Add 100 µL of Stop Solution er en ce Pu rp os eO Measure absorbance at 450 nm Fo rR ef Cat#: CY-7049 3 Version#: 140318
