Validated using a knockout cell line

Anti-FTO antibody [5-2H10] (ab92821)

Reviews (2)Q&A (2)References (19)
Western blot - Anti-FTO antibody [5-2H10] (ab92821)
  • Flow Cytometry - Anti-FTO antibody [5-2H10] (ab92821)
  • Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [5-2H10] (ab92821)
  • Western blot - Anti-FTO antibody [5-2H10] (ab92821)

Key features and details

  • Mouse monoclonal [5-2H10] to FTO
  • Suitable for: WB, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Rat, Human
  • Isotype: IgG2a

Overview

  • Product name

    Anti-FTO antibody [5-2H10]
    See all FTO primary antibodies

  • Description

    Mouse monoclonal [5-2H10] to FTO

  • Host species

    Mouse

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Sheep, Horse, Cow, Dog, Non human primates

  • Immunogen

    Synthetic peptide from Human FTO.

  • Positive control

    • WB: Rat testes nuclear lysate. ICC/IF: Neuronal progenitor cells. Flow Cyt: SH-SY5Y cells.

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab92821 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. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
ICC/IF (1)
1/100.
Flow Cyt
Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
Notes
WB
1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
ICC/IF
1/100.
Flow Cyt
Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

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 [5-2H10] (ab92821)
      Western blot - Anti-FTO antibody [5-2H10] (ab92821)

      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 - ab92821 observed at 58 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab92821 was shown to specifically recognize FTO in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when FTO knockout samples were examined. Wild-type and FTO knockout samples were subjected to SDS-PAGE. ab92821 and ab181602 (Rabbit 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry - Anti-FTO antibody [5-2H10] (ab92821)
      Flow Cytometry - Anti-FTO antibody [5-2H10] (ab92821)
      Overlay histogram showing SH-SY5Y cells stained with ab92821 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. 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 (ab92821, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [5-2H10] (ab92821)
      Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [5-2H10] (ab92821)
      ICC/IF image of ab92821 stained PC12 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab92821, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
    • Western blot - Anti-FTO antibody [5-2H10] (ab92821)
      Western blot - Anti-FTO antibody [5-2H10] (ab92821)
      Anti-FTO antibody [5-2H10] (ab92821) at 1/1000 dilution + Rat testes lysate

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

    References (19)

    Publishing research using ab92821? Please let us know so that we can cite the reference in this datasheet.

    ab92821 has been referenced in 19 publications.

    • Svobodová Kovaríková A  et al. N6-Adenosine Methylation in RNA and a Reduced m3G/TMG Level in Non-Coding RNAs Appear at Microirradiation-Induced DNA Lesions. Cells 9:N/A (2020). PubMed: 32033081
    • Tang J  et al. Global N6-methyladenosine profiling of cobalt-exposed cortex and human neuroblastoma H4 cells presents epitranscriptomics alterations in neurodegenerative disease-associated genes. Environ Pollut 266:115326 (2020). PubMed: 32827984
    • Ge L  et al. Level of N6-Methyladenosine in Peripheral Blood RNA: A Novel Predictive Biomarker for Gastric Cancer. Clin Chem 66:342-351 (2020). PubMed: 32040577
    • Xu K  et al. N6-methyladenosine demethylases Alkbh5/Fto regulate cerebral ischemia-reperfusion injury. Ther Adv Chronic Dis 11:2040622320916024 (2020). PubMed: 32426101
    • Cen S  et al. TRAF4 acts as a fate checkpoint to regulate the adipogenic differentiation of MSCs by activating PKM2. EBioMedicine 54:102722 (2020). PubMed: 32268273
    View all Publications for this product

    Customer reviews and Q&As

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    1-2 of 2 Abreviews

    Immunoprecipitation abreview for Anti-FTO antibody [5-2H10]

     Excellent
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (Cancer cell line)
    Total protein in input
    400 µg
    Immuno-precipitation step
    Protein A
    Specification
    Cancer cell line
    Read More

    Bruno Saleme

    Verified customer

    Submitted May 02 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-FTO antibody [5-2H10]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Pig Cell (Swine Kidney cells (SK))
    Permeabilization
    Yes - 0.1% Triton-X 100
    Specification
    Swine Kidney cells (SK)
    Fixative
    3% paraformaldehyde followed by methanol post fixation
    Read More

    Abcam user community

    Verified customer

    Submitted May 24 2018

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