Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-FTO antibody [EPR6895] (ab124892)

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Overview

  • Product name

    Anti-FTO antibody [EPR6895]
    See all FTO primary antibodies

  • Description

    Rabbit monoclonal [EPR6895] to FTO

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP

  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat

  • Immunogen

    Synthetic peptide within Human FTO aa 450-550. The exact sequence is proprietary.

  • Positive control

    • IHC-P: Human Breast, lung carcinoma and kidney tissues; WB: 293T, SH-SY5Y, Caco2, HEK-293 and BxPC3 lysates.

  • General notes

     

     

    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
    For more information see here.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab124892 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
WB 1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.

    • Target

    • Function

      Dioxygenase that repairs alkylated DNA and RNA by oxidative demethylation. Has highest activity towards single-stranded RNA containing 3-methyluracil, followed by single-stranded DNA containing 3-methylthymine. Has low demethylase activity towards single-stranded DNA containing 1-methyladenine or 3-methylcytosine. Has no activity towards 1-methylguanine. Has no detectable activity towards double-stranded DNA. Requires molecular oxygen, alpha-ketoglutarate and iron. Contributes to the regulation of the global metabolic rate, energy expenditure and energy homeostasis. Contributes to the regulation of body size and body fat accumulation.

    • Tissue specificity

      Ubiquitously expressed, with relatively high expression in adrenal glands and brain; especially in hypothalamus and pituitary.

    • Involvement in disease

      Defects in FTO are the cause of growth retardation developmental delay coarse facies and early death (GRDDCFED) [MIM:612938]. The disease consists of a severe children multiple congenital anomaly syndrome with death by the age of 3 years. All affected individuals had postnatal growth retardation, microcephaly, severe psychomotor delay, functional brain deficits, and characteristic facial dysmorphism. In some patients, structural brain malformations, cardiac defects, genital anomalies, and cleft palate were also observed.

    • Sequence similarities

      Belongs to the fto family.

    • Domain

      The 3D-structure of the Fe2OG dioxygenase domain is similar to that of the Fe2OG dioxygenase domain found in the bacterial DNA repair dioxygenase alkB and its mammalian orthologs, but sequence similarity is very low. As a consequence, the domain is not detected by protein signature databases.

    • Cellular localization

      Nucleus.
    • Information by UniProt

    • Database links

      • Alternative names

        • AlkB homolog 9 antibody
        • ALKBH9 antibody
        • Alpha-ketoglutarate-dependent dioxygenase FTO antibody
        • AW743446 antibody
        • Fat mass and obesity-associated protein antibody
        • FATSO, MOUSE, HOMOLOG OF antibody
        • Fto antibody
        • FTO_HUMAN antibody
        • GDFD antibody
        • KIAA1752 antibody
        • mKIAA1752 antibody
        • Protein fatso antibody
        see all

      Images

      • Western blot - Anti-FTO antibody [EPR6895] (ab124892)
        Western blot - Anti-FTO antibody [EPR6895] (ab124892)

        Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
        Lane 2: FTO knockout HAP1 whole cell lysate (20 µg)
        Lane 3: HEK293 whole cell lysate (20 µg)
        Lane 4: MOLT4 whole cell lysate (20 µg)

        Lanes 1 - 4: Merged signal (red and green). Green - ab124892 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.

        ab124892 was shown to specifically react with FTO in wild-type HAP1 cells. No band was observed when FTO knockout samples were examined. Wild-type and FTO knockout samples were subjected to SDS-PAGE. ab124892 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

      • Western blot - Anti-FTO antibody [EPR6895] (ab124892)
        Western blot - Anti-FTO antibody [EPR6895] (ab124892)
        All lanes : Anti-FTO antibody [EPR6895] (ab124892) at 1/1000 dilution (Purified)

        Lane 1 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
        Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

        Predicted band size: 58 kDa
        Observed band size: 58 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6895] (ab124892)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6895] (ab124892)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling FTO with purified ab124892 at 1:100 dilution (1.66 µg/ml). Heat mediated antigen retrieval was performed 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.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6895] (ab124892)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6895] (ab124892)

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue sections labeling FTO with purified ab124892 at 1:100 dilution (1.66 µg/ml). Heat mediated antigen retrieval was performed 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.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6895] (ab124892)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6895] (ab124892)

        ab124892, at 1/100 dilution, staining FTO in Paraffin-embedded Human Breast tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

      • Western blot - Anti-FTO antibody [EPR6895] (ab124892)
        Western blot - Anti-FTO antibody [EPR6895] (ab124892)
        All lanes : Anti-FTO antibody [EPR6895] (ab124892) at 1/1000 dilution

        Lane 1 : 293T lysate
        Lane 2 : SH-SY5Y lysate
        Lane 3 : Caco2 lysate
        Lane 4 : BxPC3 lysate

        Lysates/proteins at 10 µg per lane.

        Secondary
        All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

        Predicted band size: 58 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6895] (ab124892)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6895] (ab124892)

        ab124892, at 1/100 dilution, staining FTO in Paraffin-embedded Human kidney tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

      • Other - Anti-FTO antibody [EPR6895] (ab124892)
        Other - Anti-FTO antibody [EPR6895] (ab124892)
        Equilibrium disassociation constant (KD)
        Learn more about KD

        Click here to learn more about KD

      References

      This product has been referenced in:

      • Li Y  et al. Expression of Demethylase Genes, FTO and ALKBH1, Is Associated with Prognosis of Gastric Cancer. Dig Dis Sci N/A:N/A (2019). Read more (PubMed: 30637548) »
      • Zhuang C  et al. N6-methyladenosine demethylase FTO suppresses clear cell renal cell carcinoma through a novel FTO-PGC-1a signalling axis. J Cell Mol Med 23:2163-2173 (2019). Read more (PubMed: 30648791) »
      See all 16 Publications for this product

      Publishing research using ab124892? Please let us know so that we can cite the reference in this datasheet.

      Customer reviews and Q&As

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      Immunocytochemistry/ Immunofluorescence abreview for Anti-FTO antibody [EPR6895]

       Excellent
      Abreviews
      This product is known to not work in this application or species.
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (HepG2)
      Permeabilization
      Yes - 0.05% Triton
      Specification
      HepG2
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
      Fixative
      Paraformaldehyde
      Read More

      Fange Liu

      Verified customer

      Submitted Mar 09 2017

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