P53 Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Rabbit (1:2000) -Soak PVDF membranewith pore size 0.45 um in methanol for 30 sec -Apply Wet transfersystems at 180mA for 90 min with Wet transfer buffer.

Protocol tips
-Rabbit (1:2000) -Soak PVDF membranewith pore size 0.45 um in methanol for 30 sec -Apply Wet transfersystems at 180mA for 90 min with Wet transfer buffer.

Publication protocol

After incubation with ziyuglycoside I (5, 10, and 20 µM) for the indicated time, cells were collected and lysed in ice-cold RIPA buffer for 30 min. Protein concentration was determined by the Bradford method [31]. 50 mg of protein from each sample was separated by 15% SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Gels were transferred electrophoretically on to PVDF membranes. PVDF membranes were then blocked with 5% non-fat dry milk in the mixture of TBST (Tris-Buffered Saline and Tween-20) for 1 h, and incubated with primary antibodies at 4 °C for 12 h. Membranes were washed with TBST and incubated with corresponding secondary antibody at room temperature for 1 or 2 h. The bands on the membranes were visualized using the ECL assay kit. The density of each band was normalized to the expression of β-actin.

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Manufacturer protocol

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