PE Anti-ALDH1A1 antibody [EP1933Y]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - PE Anti-ALDH1A1 antibody [EP1933Y] (AB209437)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling ALDH1A1 with ab209437 at 1/50 dilution (Magenta).

Confocal image showing cytoplasmic staining in HepG2 cells and no staining in MCF7 cells.

Nuclear DNA was labelled with DAPI (shown in blue).

ab195887 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 488) was used to counterstain tubulin at 1/200 dilution (Green).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - PE Anti-ALDH1A1 antibody [EP1933Y] (AB209437)

Overlay histogram showing MCF7 cells stained with ab209437 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min at 22°C. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab209437, 1/500 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (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 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - PE Anti-ALDH1A1 antibody [EP1933Y] (AB209437)

Flow cytometry overlay histogram showing wild-type A549 (green line) and ALDH1A1 knockout A549 cells stained with ab209437 (magenta line). The cells were fixed with 80% methanol (5 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 (ab209437) (1x 10⁶ in 100 μl at 0.04 μg/ml (1/12500)) for 30 min at 22°C.

Isotype control antibody Rabbit IgG monoclonal Phycoerythrin Isotype Control (ab209478) was used at the same concentration and conditions as the primary antibody (wild-type A549 - black line, ALDH1A1 knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

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

This antibody gave a positive signal in wild-type A549 fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.