FITC Anti-CD90 / Thy1 antibody [MRC OX-7] (ab226)

ab226 staining CD90 / Thy1 in undifferentiated PC12 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab226 at 1/100 dilution (shown in green) and ab190573, Rabbit monoclonal to alpha Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI(shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8)

ab226 staining CD90 / Thy1 in PC12 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab226 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Flow cytometry overlay histogram showing PC12 (undifferentiated) cells stained with ab226 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction, followed by the antibody (ab226) (1x 10⁶ in 100µl at 1µg/ml (1/500)) for 30 min at 22°C.

Isotype control antibody (black line) was used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in PC12 (undifferentiated) cells fixed with 4 &#37 formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

Flow cytometry overlay histogram showing PC12 (undifferentiated) cells stained with ab226 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction, followed by the antibody (ab226) (1x 10⁶ in 100µl at 1µg/ml (1/500)) for 30 min on ice.

Isotype control antibody (black line) was  used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Overlay histogram showing PC12 cells stained with ab226 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab226, 1/500 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was Mouse IgG1 (monoclonal) FITC (ab18435) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.