Overview

  • 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.

  • Tested applications

    Suitable for: ICC/IF, WB, IP, Flow Cyt, 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: HepG2 cells. IP: HepG2 cell lysate
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
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.
Flow Cyt 1/20.

For unpurified use at 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P 1/50.

See IHC antigen retrieval protocols.

The mouse recommendation is based on the WB results. We do not guarantee IHC-P for mouse.

Target

  • 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
  • Database links

  • Alternative names

    • Acetaldehyde dehydrogenase 1 antibody
    • AHD2 antibody
    • AL1A1_HUMAN antibody
    • ALDC antibody
    • Aldehyde dehydrogenase 1 family member A1 antibody
    • Aldehyde dehydrogenase 1 soluble antibody
    • Aldehyde dehydrogenase 1A1 antibody
    • Aldehyde dehydrogenase antibody
    • Aldehyde dehydrogenase cytosolic antibody
    • Aldehyde dehydrogenase family 1 member A1 antibody
    • Aldehyde dehydrogenase liver cytosolic antibody
    • ALDH 1 antibody
    • ALDH 1A1 antibody
    • ALDH class 1 antibody
    • ALDH, liver cytosolic antibody
    • ALDH-E1 antibody
    • ALDH1 A1 antibody
    • ALDH1 antibody
    • ALDH11 antibody
    • ALDH1A1 antibody
    • ALHDII antibody
    • cytosolic antibody
    • epididymis luminal protein 12 antibody
    • epididymis luminal protein 9 antibody
    • epididymis secretory sperm binding protein Li 53e antibody
    • HEL-S-53e antibody
    • MGC2318 antibody
    • PUMB1 antibody
    • RALDH 1 antibody
    • RalDH1 antibody
    • Retinal dehydrogenase 1 antibody
    see all

Images

  • 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

  • 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 minute


    Blocking/Diluting buffer 5% NFDM/TBST

  • 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.
  • 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.

  • 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.

  • 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).

  • 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.

  • Sections of the proximal (top) and distal (bottom) mouse colon were assayed for protein expression of ALDH1A1.

    Signals yielded by immunohistochemical staining (red) are shown in merged images with DAPI staining (blue). Representative data are shown for three independent experiments. Arrowheads show positive cells for ALDH1A1 in the distal colon. Scale bars, 50 μm.

    Colonic tissues were fixed overnight at 4°C in 4% paraformaldehyde, sequentially dehydrated in 10, 15 and 20% sucrose in PBS, and embedded in OCT compound. Colonic organoids were fixed together with surrounding collagen type I gel and processed in the same manner. Frozen sections of 8-μm thickness were subjected to immunohistochemistry using ab52492 (unpurified). 

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

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

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

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

  • 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.

References

This product has been referenced in:

  • Zhang C & Wang Y Metformin attenuates cells stemness and epithelial-mesenchymal transition in colorectal cancer cells by inhibiting the Wnt3a/ß-catenin pathway. Mol Med Rep 19:1203-1209 (2019). Read more (PubMed: 30569135) »
  • Wu L  et al. Disulfiram and BKM120 in Combination with Chemotherapy Impede Tumor Progression and Delay Tumor Recurrence in Tumor Initiating Cell-Rich TNBC. Sci Rep 9:236 (2019). Read more (PubMed: 30659204) »
See all 93 Publications for this product

Customer reviews and Q&As

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1-10 of 16 Abreviews

Application
Western blot
Loading amount
50 µg
Gel Running Conditions
Reduced Denaturing (12% acrylamide gel)
Sample
Human Tissue lysate - whole (Brain)
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 24 2013

Application
Western blot
Sample
Mouse Cell lysate - whole cell (fibroblast)
Loading amount
30 µg
Specification
fibroblast
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 23 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Specification
Liver
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 37°C

Marta Canamero

Verified customer

Submitted Aug 21 2008

Application
Flow Cytometry
Sample
Human Cell (single cells of xenograft human breast cancer cell)
Permeabilization
No
Gating Strategy
0.5-1%
Specification
single cells of xenograft human breast cancer cell
Preparation
Cell harvesting/tissue preparation method: Dissociated cells from FFFE xenograft tissues
Sample buffer: 1% BSA

Abcam user community

Verified customer

Submitted Sep 17 2019

Application
Western blot
Sample
Human Cell lysate - whole cell (CWR-R1 Prostate cancer cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
CWR-R1 Prostate cancer cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 05 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (PC3 Prostate cancer cells)
Permeabilization
Yes - 0.1% Triton X-100
Specification
PC3 Prostate cancer cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Dr. Dimitra Kalamida

Verified customer

Submitted Mar 21 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Pancreatic Cancer)
Permeabilization
No
Specification
Pancreatic Cancer
Blocking step
(agent) for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Sep 25 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Pancreatic)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
10 µg
Specification
Pancreatic
Blocking step
NET Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Sep 25 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (Melanoma cancer cell line (A375))
Gel Running Conditions
Non-reduced Denaturing (4-20 % gel)
Loading amount
10 µg
Specification
Melanoma cancer cell line (A375)
Blocking step
Odyssey Block buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C

Miss. Rebecca Atkinson-Dell

Verified customer

Submitted Aug 08 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (10%)
Loading amount
15 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 19 2015

1-10 of 16 Abreviews

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