Alexa Fluor® 488 Anti-Cytokeratin 17 抗体 [EP1623] - Cytoskeleton Marker (ab185032)

Overlay histogram showing HeLa cells stained with ab185032 (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 (ab185032, 1/500 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 488 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 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

This antibody gave a positive signal in HeLa fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Immunohistochemistry analysis of paraffin-embedded human tonsil tissue sections labelling Cytokeratin 17 with ab185032 at 1/100 dilution. The section was incubated with ab185032 for 60 mins at room temperature (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount^®. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 40 mins.
Membrane staining on human tonsil tissue. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Negative Control: The negative control is PBS.

ab185032 staining Cytokeratin 17 in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab185032 at 1/50 dilution overnight at +4°C (shown in green). AlexaFluor^®350 WGA was used at a 1/200 dilution and incubated for 1h with the cells, to label plasma membranes (shown in blue). Nuclear DNA was labelled in red with 1.25 μM DRAQ5™ (ab108410).

Immunohistochemistry analysis of paraffin-embedded human bladder cancer tissue sections labelling Cytokeratin 17 with ab185032 at 1/100 dilution. The section was incubated with ab185032 for 60 mins at room temperature (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount^®. Antigen retrieval was heat mediated with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 40 mins.
Membrane staining on human bladder cancer tissue. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Negative Control: The negative control is PBS.