Alexa Fluor® 488 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (AB184818)

ab184818 staining Hexokinase 1 in HeLa cells. The cells were fixed with 4% PFA (10min) 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 ab184818 at a working dilution of 1 in 100 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).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (AB184818)

Overlay histogram showing HeLa cells stained with ab184818 (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 (ab184818, 1/50 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 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Anti-Hexokinase 1 antibody [EPR10134(B)] - Mitochondrial Outer Membrane Marker (AB184818)

Immunofluorescent analysis of 4% PFA-fixed 0.1% Triton X-100 permeabilized HEK-293 WT and HEK-293-HK1 KO cells labelling Hexokinase 1 with ab184818 at 0.2 μg/ml concentration (Green). Cells were counterstained with ab190573 Anti-Tubulin (Alexa Fluor® 647) at 1 : 250 (2 μg/ml) (Magenta). The nuclear counterstain was DAPI (Blue).