Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling ALDH1A1 with purified ab52492 at 1/50 dilution (3.54 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

ab52492 (purified) at 1/20 dilution (2ug) immunoprecipitating ALDH1A1 in HepG2 whole cell lysates.
Lane 1: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates 10ug
Lane 2 (+): ab52492 & HepG2 whole cell lysates
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52492 in HepG2 whole cell lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling ALDH1A1 with ab52492 (purified) at 1/500 dilution (4 μg/ml).

Cells were fixed in 100% methanol. ab150077, an AlexaFluor^®488 Goat anti-Rabbit secondary antibody was used at 1/1000 dilution (2 μg/ml). ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain at 1/200 dilution (2.5 μg/ml). DAPI was used as nuclear counterstain.

Confocal image showing cytoplasmic staining on HepG2 cell line.

Negative control: No staining on MCF-7 cell line.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

Clone EP1933Y (ab215996) has been successfully conjugated by Abcam. This image was generated using Anti-ALDH1A1 antibody [EP1933Y] (PE). Please refer to ab209437 for protocol details.

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.

Clone EP1933Y (ab215996) has been successfully conjugated by Abcam. This image was generated using Anti-ALDH1A1 antibody [EP1933Y] (Alexa Fluor® 647). Please refer to ab195255 for protocol details.

ab195255 staining ALDH1A1 in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes 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 ab195255 at a working dilution of 1 in 50 (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Clone EP1933Y (ab215996) has been successfully conjugated by Abcam. This image was generated using Anti-ALDH1A1 antibody [EP1933Y] (Alexa Fluor® 488). Please refer to ab195254 for protocol details.

ab195254 staining ALDH1A1 in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised in 0.1% Triton X-100 for 5 minutes 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 ab195254 at a working dilution of 1 in 100 (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor^® 594, shown in red) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ALDH1A1 with purified ab52492 at 1/20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

Tumor tissues of primary invasive ductal carcinomas of the breast were obtained from 192 female patients with stage IIB and III prior to pre-operative neoadjuvant chemotherapy.

(progressive or stable disease, PD/SD)

(partial or complete remission, PR/CR)

The level of ALDH1 was tested by immunohistochemistry staining in paraffin-embedded tissue sections. Rabbit monoclonal ALDH1A1 antibody (ab52492, unpurified, Abcam) used at a 1:100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

Overlay histogram showing HepG2 (Human liver hepatocellular carcinoma cell line) cells stained with ab52492 (unpurified) (red line).

The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab52492, 1/1000 dilution) for 30 minutes at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabeled 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 data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

Immunohistochemical analysis of paraffin-embedded human liver tissue sections labeling ALDH1A1 with ab52492 (unpurified) at 1/100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded normal human brain tissue sections labeling ALDH1A1 with ab52492 (unpurified).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded normal human lung tissue sections labeling ALDH1A1 with ab52492 (unpurified).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).

Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded normal human kidney tissue sections labeling ALDH1A1 with ab52492 (unpurified).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52492).