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    fto-antibody-epr6894-ab126605.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-FTO antibody [EPR6894] (ab126605)

  • Datasheet
Reviews (3) Submit a question References (14)

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Western blot - Anti-FTO antibody [EPR6894] (ab126605)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (ab126605)
  • Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)
  • Western blot - Anti-FTO antibody [EPR6894] (ab126605)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (ab126605)
  • Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)
  • Western blot - Anti-FTO antibody [EPR6894] (ab126605)
  • Anti-FTO antibody [EPR6894] (ab126605)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6894] to FTO
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free

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Recombinant Human FTO protein (ab109039)
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Alexa Fluor® 488 Anti-FTO antibody [EPR6894] (ab207665)

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Overview

  • Product name

    Anti-FTO antibody [EPR6894]
    See all FTO primary antibodies
  • Description

    Rabbit monoclonal [EPR6894] to FTO
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human FTO (N terminal). The exact sequence is proprietary.

  • Positive control

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

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 59% PBS, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR6894
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Metabolism
    • Cardiovascular
    • Atherosclerosis
    • Lipoprotein metabolism
    • Cardiovascular
    • Atherosclerosis
    • Lipid transport
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipoprotein metabolism
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Obesity
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease
    • Metabolism
    • Types of disease
    • Metabolic disorders

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-FTO antibody [EPR6894] (ab207665)
    • Anti-FTO antibody [EPR6894] - BSA and Azide free (ab240021)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human FTO protein (ab109039)

Applications

The Abpromise guarantee

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)
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

See IHC antigen retrieval protocols.

ICC/IF (1)
1/250.

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

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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

See IHC antigen retrieval protocols.

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.
  • Target information above from: UniProt accession Q9C0B1 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 79068 Human
    • Omim: 610966 Human
    • SwissProt: Q9C0B1 Human
    • Unigene: 528833 Human
    • 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 [EPR6894] (ab126605)
      Western blot - Anti-FTO antibody [EPR6894] (ab126605)

      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 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. ab126605 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (ab126605)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (ab126605)

      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)
      Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)

      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.

    • Western blot - Anti-FTO antibody [EPR6894] (ab126605)
      Western blot - Anti-FTO antibody [EPR6894] (ab126605)
      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
      All lanes : 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (ab126605)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (ab126605)

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

    • Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)
      Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)

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

    • Western blot - Anti-FTO antibody [EPR6894] (ab126605)
      Western blot - Anti-FTO antibody [EPR6894] (ab126605)
      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
      All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

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

    • Anti-FTO antibody [EPR6894] (ab126605)
      Anti-FTO antibody [EPR6894] (ab126605)

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (14)

    Publishing research using ab126605? Please let us know so that we can cite the reference in this datasheet.

    ab126605 has been referenced in 14 publications.

    • Zhang Z  et al. Kinase GSK3ß functions as a suppressor in colorectal carcinoma through the FTO-mediated MZF1/c-Myc axis. J Cell Mol Med 25:2655-2665 (2021). PubMed: 33533172
    • Qin B  et al. Long non-coding RNA CASC15 facilitates esophageal squamous cell carcinoma tumorigenesis via decreasing SIM2 stability via FTO-mediated demethylation. Oncol Rep 45:1059-1071 (2021). PubMed: 33650646
    • Zepecki JP  et al. miRNA-mediated loss of m6A increases nascent translation in glioblastoma. PLoS Genet 17:e1009086 (2021). PubMed: 33684100
    • Tsuruta N  et al. RNA N6-methyladenosine demethylase FTO regulates PD-L1 expression in colon cancer cells. Biochem Biophys Res Commun 530:235-239 (2020). PubMed: 32828292
    • Zhao J  et al. Zbtb7b suppresses aseptic inflammation by regulating m6A modification of IL6 mRNA. Biochem Biophys Res Commun 530:336-341 (2020). PubMed: 32828308
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-FTO antibody [EPR6894]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (Skin)
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    Submitted May 10 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-FTO antibody [EPR6894]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (lung carcinoma)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mm Sodium citric, PH6.0
    Permeabilization
    No
    Specification
    lung carcinoma
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 19 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-FTO antibody [EPR6894]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Fange Liu

    Verified customer

    Submitted Feb 08 2017

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