Anti-Human p21 monoclonal antibody

Western blot analysis of p21 Waf1/Cip1 was performed by loading HUVEC and HeLa cell lysates on a SDS-PAGE gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% nonfat dry milk in TBST for 1 hour at room temperature. The membrane was probed with a p21 Waf1/Cip1 monoclonal antibody at a dilution of 1:1000 overnight at 4C, followed by an anti-rabbit IgG-HRP secondary antibody. Detection was performed using a chemiluminescent substrate.

Flow cytometric analysis of p21 Waf1/Cip1 on HeLa (red histogram) and MCF7 cells (blue histogram). Cells were harvested, fixed with formaldehyde, permeabilized with methanol, and incubated with a p21 Waf1/Cip1 monoclonal antibody at a dilution of 1:200 for 1 hour at room temperature. Cells were stained with a fluorescently-conjugated anti-rabbit IgG secondary antibody and analyzed on a flow cytometer.