Overview

  • Product name
    Anti-FTO antibody [EPR6894]
    See all FTO primary antibodies
  • Description
    Rabbit monoclonal [EPR6894] to FTO
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human FTO (N terminal).

  • Positive control
    • MOLT4, 293T, SH SY5Y, BxPC3 and Caco 2 cell lysates; Human adrenal gland tissue; BxPC3 cells.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these 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 ab126605 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 1/10000 - 1/20000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).

For unpurified, use 1/1000 - 1/10000.

IHC-P 1/500.

For unpurified, use 1/50 - 1/100.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/250.

For unpurified, use 1/50 - 1/100.

  • Application notes
    Is unsuitable for Flow Cyt.
  • 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



    • Predicted band size : 58 kDa

      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 - ab126605 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab126605 was shown to specifically react with FTO when FTO knockout samples were used. Wild-type and FTO knockout samples were subjected to SDS-PAGE. ab126605 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemical staining of paraffin embedded human hepatocellular carcinoma with purified ab126605 at a working dilution of 1 in 500. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • Immunofluorescence staining of BxPC-3 cells with purified ab126605 at a working dilution of 1 in 250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit, used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab126605 was used at a dilution of 1/200 followed by an Alexa Fluor® 488 goat anti-mouse antibody at a dilution of 1/500.

    • All lanes : Anti-FTO antibody [EPR6894] (ab126605) at 1/10000 dilution (purified)

      Lane 1 : Molt-4 cell lysate
      Lane 2 : HEK293 cell lysate
      Lane 3 : BxPC-3 cell lysate
      Lane 4 : Caco-2 cell lysate
      Lane 5 : SH-SY5Y cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

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

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical staining of FTO in paraffin embedded human adrenal gland tissue using unpurified ab126605 at a 1/50 dilution.

    • Unpurified ab126605 at a 1/50 dilution staining FTO in BxPC3 cells by immunofluorescence.

    • All lanes : Anti-FTO antibody [EPR6894] (ab126605) at 1/1000 dilution (unpurified)

      Lane 1 : MOLT4 cell lysate
      Lane 2 : 293T cell lysate
      Lane 3 : SH SY5Y cell lysate
      Lane 4 : BxPC3 cell lysate
      Lane 5 : Caco 2 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat anti-Rabbit HRP at 1/2000 dilution

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

    References

    This product has been referenced in:
    • Aas A  et al. Nucleocytoplasmic Shuttling of FTO Does Not Affect Starvation-Induced Autophagy. PLoS One 12:e0168182 (2017). WB, ICC/IF ; Mouse, Human . Read more (PubMed: 28288181) »

    See 1 Publication for this product

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Mel-624)
    Permeabilization
    Yes - 0.05% Triton
    Specification
    Mel-624
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Fange Liu

    Verified customer

    Submitted Feb 08 2017

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

    Sign up