PE Anti-Glucose Transporter GLUT1 antibody [EPR3915]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - PE Anti-Glucose Transporter GLUT1 antibody [EPR3915] (AB209449)

Overlay histogram showing HepG2 cells stained with ab209449 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol (-20°C) for 30 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab209449, 1/2500 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycorythrin (ab209478) 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 20 mW Solid State Blue Laser (488nm) and 585/40 bandpass filter.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - PE Anti-Glucose Transporter GLUT1 antibody [EPR3915] (AB209449)

ab209449 staining Glucose Transporter GLUT1 in HepG2 cells. The cells were fixed with 4% formaldehyde (10 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 ab209449 at 1/500 dilution (Pseudocolored in green) and ab195884, Rat monoclonal to 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).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - PE Anti-Glucose Transporter GLUT1 antibody [EPR3915] (AB209449)

Immunofluorescent analysis of 4% PFA-fixed 0.1% Triton X-100 permeabilized A549 WT and A549-SLC2A1 KO cells labelling Glucose Transporter GLUT1 with ab209449 at 1 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).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - PE Anti-Glucose Transporter GLUT1 antibody [EPR3915] (AB209449)

Flow cytometry overlay histogram showing wild-type A-549 (green line) and SLC2A1 knockout A-549 stained with ab209449 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab209449) (1x 10⁶ in 100μl at 0.008 μg/ml (1/62500)) for 30min at 22°C.

Isotype control antibody was Rabbit IgG (monoclonal) Phycoerythrin Isotype Control (ab209478) in A-549 WT cells (black line) and A-549-SLC2A1 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Yellow/Green laser (561nm) and 585/42 bandpass filter.