Anti-ALDH1A1 抗体 [EP1933Y] - C-terminal

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

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

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (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 (unpurified ab52492, Abcam) used at a 1 : 100 dilution.

Image from Gong et al PLoS One. 2010 Dec 20;5(12):e15630. doi: 10.1371/journal.pone.0015630. Fig 1.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

Overlay histogram showingHepG2 (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 Fluorr^® 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ALDH1A1 with purified ab52492 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

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

• IP

Unknown

Immunoprecipitation - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

ab52492 (purified) at 1/20 dilution (2μg) immunoprecipitating ALDH1A1 in HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates.
Lane 1 : HepG2 whole cell lysates 10μg.
Lane 2 : ab52492 IP in 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.

All lanes:

Immunoprecipitation - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492)

Predicted band size: 54 kDa

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• WB

Lab

Western blot - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

All lanes:

Western blot - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492) at 1/1000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

Lane 2:

Human small intestine lysates

Lane 3:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 4:

MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 5:

Mouse liver lysates at 20 µg

Lane 6:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 54 kDa

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

Western blot : Anti-ALDH1A1 antibody [EP1933Y] (ab52492) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52492 was shown to bind specifically to ALDH1A1. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in ALDH1A1 knockout cell line. To generate this image, wild-type and ALDH1A1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

ALDH1A1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human ALDH1A1 knockout A549 cell line (<a href='/products/cell-lines/human-aldh1a1-knockout-a549-cell-line-ab261864'>ab261864</a>)

Lane 2:

Western blot - Human ALDH1A1 knockout A549 cell lysate (<a href='/products/cell-lysates/human-aldh1a1-knockout-a549-cell-lysate-ab261673'>ab261673</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

false

• WB

Lab

Western blot - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (AB52492)

Blocking/Diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492) at 1/1000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Human small intestine tissue lysate at 15 µg

Lane 3:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 4:

MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 5:

Mouse liver tissue lysate at 15 µg

Lane 6:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 15 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 54 kDa

Observed band size: 55 kDa

false

Exposure time: 1min