Alexa Fluor® 647 Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - Neuronal Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - Neuronal Marker (AB199829)

ab199829 staining ENO1 + ENO2 + ENO3 in U87MG cells. The cells were fixed with 100% methanol (5min), 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 ab199829 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 2μg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal high-content analyser (Perkin Elmer, Operetta).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - Neuronal Marker (AB199829)

Overlay histogram showing U-87MG cells stained with ab199829 (red line). The cells were fixed with 4% formaldehyde (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 (ab199829, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG [EPR25A] Alexa Fluor® 647 (ab199093) 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 solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter. This antibody gave a positive signal in U-87MG cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.