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
  • Tested applications
    Suitable for: ICC/IF, WB, IP, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Does not react with: Rat
  • Immunogen

    Synthetic peptide within Human ALDH1A1 aa 300-400 (C terminal). The exact sequence is proprietary.

  • 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. ICC/IF: HepG2 cells. Flow Cyt: HepG2 cells.
  • General notes

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

    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 1/30.
Flow Cyt 1/1000.

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

IHC-P Use at an assay dependent concentration.

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

  • 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, 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. 

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

  • All lanes : Anti-ALDH1A1 antibody [EP1933Y] - C-terminal (ab52492) at 1/1000 dilution

    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

  • Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling ALDH1A1 with ab52492 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.

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

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

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

  • Overlay histogram showing HepG2 (Human liver hepatocellular carcinoma cell line) cells stained with ab52492 (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:
  • Boulding T  et al. LSD1 activation promotes inducible EMT programs and modulates the tumour microenvironment in breast cancer. Sci Rep 8:73 (2018). Read more (PubMed: 29311580) »
  • Shi Y  et al. Nuclear EGFR-PKM2 axis induces cancer stem cell-like characteristics in irradiation-resistant cells. Cancer Lett 422:81-93 (2018). WB, ChIP . Read more (PubMed: 29477380) »
See all 78 Publications for this product

Customer reviews and Q&As

1-6 of 6 Q&A

Answer

Thank you for submitting an Abreview for ab52492. As you may have noticed, your review has now been published on our website.

Since you obtained poor results using the antibody in an untested application, we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.

- Could you please let me knowwhat was the exact problem you have encountered?
- Have you tried the antibody on another tested application?

I look forward to hear back from you.

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Question
Answer

Vielen Dank für Ihren Anruf.

Ich freue mich Ihnen nun bestätigen zu können, dass die Konzentration 0.130 mg/mL ist. (von Lot 1315178).

Ich würde Sie außerdem noch gerne darauf hinweisen, dass ich für eine effiziente Konjugierung mit ab102908 (EasyLink PerCP Conjugation Kit (3 x 10µg PerCP)) eine Aufreinigung von ab52492 dringend empfehle.

ab52492 ist Zellkulturüberstand, der viele andere Proteine enthält. Das Easylink Kit wird alle zugänglichen Amine mit PerCP konjugieren, und somit auch die anderen Proteine in der Antikörperlösung.

Ich kann Ihnen das folgende Kit empfehlen:

ab109207 (Antibody TCS Purification Kit)

https://www.abcam.com/index.html?datasheet=109207 (or use the following: https://www.abcam.com/index.html?datasheet=109207).

Ich hoffe, diese Information was hilfreich und wünsche Ihnen viel Erfolg für Ihre Experimente.

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Answer

Thank you for contacting Abcam.

Based on the information we have for ab52492, we would recommend that you use heat mediated antigen retrieval with sodium citrate. I have attached our Abcams protocol booklet that contains information about this methods and also contains recipes for the solutions aswell.

If there is anything else I can help you with, please let me know.

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Question
Answer

Thank you for contacting Abcam.



The immunogen shares 88% identity with canine Aldh1a1.

Please let me know if there is anything else I can help you with.

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Answer

Thank you for contacting Abcam regarding ab52492.


I am sorry that you are experiencing some difficulties detecting the correct molecular weight band in WB with this antibody. I have reviewed the protocol information you provided and I am not sure that any changes to this would improve your results. Have you completely denatured and reduced the protein?I would recommend boiling for10 minutes and be sure you have included DTT or beta-mercaptoethanol in the sample buffer.


Another explanationis that this protein is subject to post-translational modifications, acetylation in particular at 10 different residues, resulting in a shift of molecular weight.

Unfortunately a blocking peptide is not available, but I can provide the protein sequence if you were interested in synthesizing your own.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions or concerns.

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Question
Answer

Thank you for your phone call today. I contacted the lab and was given the protocol used for IHC testing. Antigen retrieval was performed using pH 6 citrate buffer for 20-30 minutes in a heated rice cooker. The exact dilution of this antibody for IHC is variable, but I recommend starting with 1:100 dilution. I have attached the full protocol to this email. I hope this information is helpful, but please let me know if you have further questions.

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