Recombinant Anti-FTO antibody [EPR6894] (ab126605)
![Western blot - Anti-FTO antibody [EPR6894] (ab126605)](https://www.abcam.com/ps/products/126/ab126605/Images/ab126605-280296-anti-fto-antibody-epr6894-western-blot.jpg "Western blot - 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
Overview
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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.
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Positive control
- MOLT4, 293T, SH SY5Y, BxPC3 and Caco 2 cell lysates; Human adrenal gland tissue; BxPC3 cells.
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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.
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
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
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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
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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. |
| 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. |
|
ICC/IF
1/250. For unpurified, use 1/50 - 1/100. |
Target
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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
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Database links
- Entrez Gene: 79068 Human
- Omim: 610966 Human
- SwissProt: Q9C0B1 Human
- Unigene: 528833 Human
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Alternative names
- AlkB homolog 9 antibody
- ALKBH9 antibody
- Alpha-ketoglutarate-dependent dioxygenase FTO antibody
see all
Images
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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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (ab126605)](https://www.abcam.com/ps/products/126/ab126605/Images/ab126605-241015-anti-fto-antibody-epr6894-immunohistochemistry.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)](https://www.abcam.com/ps/products/126/ab126605/Images/ab126605-241014-anti-fto-antibody-epr6894-immunofluorescence.jpg)
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.
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![Western blot - Anti-FTO antibody [EPR6894] (ab126605)](https://www.abcam.com/ps/products/126/ab126605/Images/ab126605-241012-anti-fto-antibody-epr6894-western-blot.jpg)
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 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FTO antibody [EPR6894] (ab126605)](https://www.abcam.com/ps/products/126/ab126605/Images/ab126605-140572-anti-fto-antibody-epr6894-immunohistochemistry.jpg)
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.
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![Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)](https://www.abcam.com/ps/products/126/ab126605/Images/ab126605-140571-anti-fto-antibody-epr6894-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-FTO antibody [EPR6894] (ab126605)Unpurified ab126605 at a 1/50 dilution staining FTO in BxPC3 cells by immunofluorescence.
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![Western blot - Anti-FTO antibody [EPR6894] (ab126605)](https://www.abcam.com/ps/products/126/ab126605/Images/ab126605-140570-anti-fto-antibody-epr6894-western-blot.jpg)
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)](https://www.abcam.com/ps/products/126/ab126605/Images/ab126605-9-benefits-of-recombinant-antibodies.png)
Anti-FTO antibody [EPR6894] (ab126605)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab126605 has been referenced in 8 publications.
- 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
- Yang S et al. m6A mRNA demethylase FTO regulates melanoma tumorigenicity and response to anti-PD-1 blockade. Nat Commun 10:2782 (2019). PubMed: 31239444
- Baquero-Perez B et al. The Tudor SND1 protein is an m6A RNA reader essential for replication of Kaposi's sarcoma-associated herpesvirus. Elife 8:N/A (2019). WB ; Human . PubMed: 31647415
- Koh CWQ et al. Atlas of quantitative single-base-resolution N6-methyl-adenine methylomes. Nat Commun 10:5636 (2019). PubMed: 31822664
- Wang X et al. mRNA m6A plays opposite role in regulating UCP2 and PNPLA2 protein expression in adipocytes. Int J Obes (Lond) 42:1912-1924 (2018). PubMed: 29487348
- Aas A et al. Nucleocytoplasmic Shuttling of FTO Does Not Affect Starvation-Induced Autophagy. PLoS One 12:e0168182 (2017). WB, ICC/IF ; Mouse, Human . PubMed: 28288181
- Xiang Y et al. RNA m6A methylation regulates the ultraviolet-induced DNA damage response. Nature 543:573-576 (2017). PubMed: 28297716