Recombinant Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1933Y] to ALDH1A1 - C-terminal
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-ALDH1A1 antibody [EP1933Y] - C-terminal
See all ALDH1A1 primary antibodies -
Description
Rabbit monoclonal [EP1933Y] to ALDH1A1 - C-terminal -
Host species
Rabbit -
Specificity
The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
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Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide within Human ALDH1A1 aa 300-400 (C terminal). The exact sequence is proprietary.
Database link: P00352 -
Positive control
- WB: HepG2 and NIH/3T3 cell lysate; Human small intestine and mouse liver tissue lysate. IHC: Human liver, kidney, lung and brain tissue, human breast carcinoma. ICC/IF: HepG2 cells. Flow Cyt (intra): HepG2 cells. IP: HepG2 cell lysate
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General notes
The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1933Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-ALDH1A1 antibody [EP1933Y] (ab195254)
- Alexa Fluor® 647 Anti-ALDH1A1 antibody [EP1933Y] (ab195255)
- HRP Anti-ALDH1A1 antibody [EP1933Y] (ab195517)
- Alexa Fluor® 594 Anti-ALDH1A1 antibody [EP1933Y] (ab206884)
- PE Anti-ALDH1A1 antibody [EP1933Y] (ab209437)
- Anti-ALDH1A1 antibody [EP1933Y] - BSA and Azide free (ab215996)
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab52492 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
Flow Cyt (Intra) |
1/20.
For unpurified use at 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (3) |
1/100 - 1/250.
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WB | (11) |
1/500 - 1/2000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
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IP |
1/20 - 1/30.
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IHC-P | (5) |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse. |
Notes |
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Flow Cyt (Intra)
1/20. For unpurified use at 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/250. |
WB
1/500 - 1/2000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa). |
IP
1/20 - 1/30. |
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse. |
Target
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Function
Binds free retinal and cellular retinol-binding protein-bound retinal. Can convert/oxidize retinaldehyde to retinoic acid. -
Pathway
Cofactor metabolism; retinol metabolism. -
Sequence similarities
Belongs to the aldehyde dehydrogenase family. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 216 Human
- Entrez Gene: 11668 Mouse
- Omim: 100640 Human
- SwissProt: P00352 Human
- SwissProt: P24549 Mouse
- Unigene: 76392 Human
- Unigene: 250866 Mouse
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Alternative names
- Acetaldehyde dehydrogenase 1 antibody
- AHD2 antibody
- AL1A1_HUMAN antibody
see all
Images
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All lanes : Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : ALDH1A1 knockout A549 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDaWestern 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.
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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.
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All lanes : Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492) at 1/1000 dilution (Purified)
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 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDa -
Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded normal human kidney tissue sections labeling ALDH1A1 with ab52492 (unpurified).
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All lanes : Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492) at 1/1000 dilution (unpurified)
Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : Human small intestine tissue lysate
Lane 3 : PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 5 : Mouse liver tissue lysate
Lane 6 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 55 kDa
Observed band size: 55 kDa
Exposure time: 1 minuteBlocking/Diluting buffer 5% NFDM/TBST
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Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded normal human brain tissue sections labeling ALDH1A1 with ab52492 (unpurified).
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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).
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Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded normal human lung tissue sections labeling ALDH1A1 with ab52492 (unpurified).
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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.
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Immunohistochemical analysis of paraffin-embedded human liver tissue sections labeling ALDH1A1 with ab52492 (unpurified) at 1/100 dilution.
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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. -
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.
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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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (170)
ab52492 has been referenced in 170 publications.
- Laraba L et al. Inhibition of YAP/TAZ-driven TEAD activity prevents growth of NF2-null schwannoma and meningioma. Brain 146:1697-1713 (2023). WB, IF ; Human . PubMed: 36148553
- Huttala O et al. Decellularized In Vitro Capillaries for Studies of Metastatic Tendency and Selection of Treatment. Biomedicines 10:N/A (2022). PubMed: 35203480
- Zhang S et al. GIT1 protects against breast cancer growth through negative regulation of Notch. Nat Commun 13:1537 (2022). PubMed: 35318302
- Caminear MW et al. Disulfiram Transcends ALDH Inhibitory Activity When Targeting Ovarian Cancer Tumor-Initiating Cells. Front Oncol 12:762820 (2022). PubMed: 35372040
- Ozaki A et al. High Expression of p62 and ALDH1A3 Is Associated With Poor Prognosis in Luminal B Breast Cancer. Anticancer Res 42:3299-3312 (2022). PubMed: 35790283