Effects of FZE on apoptotic ratio and apoptotic factors in RSC96 cells.

Effects of FZE on translocation of CytoC and the levels of caspase9 and caspase3. The cells were fixed with 4% paraformaldehyde for 15 minutes at 20°C, permeated with 0.3% triton prior to being blocked in 1% BSA+2% normal goat serum for 30 min at 20°C. Samples were then incubated with primary antibody overnight at 4°C in PBS containing. ab96899 diluted at 1∶200 was used as the secondary antibody. Cell nucleus were counterstained with DAPI and showed blue. Mitochondria were labeled by Mito tracker and showed red.

ICC/IF image of (ab3280) stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

The cells were fixed in 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3280, 5 µg/ml) overnight at +4°C.

The secondary antibody (green) was DyLight^® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1 h. Alexa Fluor^® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

Emission spectra of DyLight^® fluorochromes available in our catalog.
Line colors represent the approximate visible colors of the wavelength maxima.

Overlay histogram showing A431 cells stained with unpurified ab124962 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab124962, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

ab96379 staining MEK1 (phospho S298) in SK-N-SH cells treated with CNQX (ab120017), by ICC/IF. Decrease in MEK1 (phospho S298) expression correlates with increased concentration of CNQX, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120017 (CNQX) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab96379 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.

Overlay histogram showing Raji cells stained with ab52922 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52922, 1/100) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.