Recombinant Anti-FTO antibody [EPR24440-12] (ab280081)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24440-12] to FTO
- Suitable for: WB, Flow Cyt (Intra), ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-FTO antibody [EPR24440-12]
See all FTO primary antibodies -
Description
Rabbit monoclonal [EPR24440-12] to FTO -
Host species
Rabbit -
Tested applications
Suitable for: WB, Flow Cyt (Intra), ICC/IFmore details
Unsuitable for: IHC-P or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HAP1, HEK-293, NIH/3T3, C2C12, HeLa, PC-12, L6, Human lung, Human kidney, Human skeletal muscle, Mouse brain and Mouse heart, Rat brain lysates. ICC/IF: NIH/3T3 cells. Flow Cyt(intra): NIH/3T3 cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24440-12 -
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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab280081 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 |
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WB |
1/1000. Predicted molecular weight: 58 kDa.
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Flow Cyt (Intra) |
1/500.
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ICC/IF |
1/50.
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Notes |
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WB
1/1000. Predicted molecular weight: 58 kDa. |
Flow Cyt (Intra)
1/500. |
ICC/IF
1/50. |
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
- Entrez Gene: 26383 Mouse
- Entrez Gene: 291905 Rat
- Omim: 610966 Human
- SwissProt: Q9C0B1 Human
- SwissProt: Q8BGW1 Mouse
- SwissProt: Q2A121 Rat
- Unigene: 528833 Human
see all -
Alternative names
- AlkB homolog 9 antibody
- ALKBH9 antibody
- Alpha-ketoglutarate-dependent dioxygenase FTO antibody
see all
Images
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All lanes : Anti-FTO antibody [EPR24440-12] (ab280081) at 1/1000 dilution
Lane 1 : Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate
Lane 2 : FTO knockout HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate
Lane 3 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Predicted band size: 58 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 3% NFDM/TBST
Lanes 1-3: Merged signal (red and green). Green - ab280081 observed at 60 kDa. Red - loading control ab8245 observed at 36 kDa.
ab280081 Anti-FTO antibody [EPR24440-12] was shown to specifically react with FTO in wild-type HAP1 cells. Loss of signal was observed when the knockout cell line was used. Wild-type and FTO knockout samples were subjected to SDS-PAGE. ab280081 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4? overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-FTO antibody [EPR24440-12] (ab280081) at 1/1000 dilution
Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 2 : C2C12 (mouse myoblasts myoblast) whole cell lysate
Lane 3 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 5 : HAP1 (human chronic myelogenous leukemia near-haploid cell line) whole cell lysate
Lane 6 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lane 7 : L6 (rat skeletal muscle myoblast) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 58 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: Lanes 1-2:15 seconds; Lanes 3-5: 6 seconds; Lanes:6-7:15 seconds.
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All lanes : Anti-FTO antibody [EPR24440-12] (ab280081) at 1/1000 dilution
Lane 1 : Human lung tissue lysate 20
Lane 2 : Human kidney tissue lysate
Lane 3 : Human skeletal muscle tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution (VeriBlot for IP secondary antibody(HRP))
Predicted band size: 58 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: Lanes 1-2: 15 seconds;Lane 3: 48 seconds.
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All lanes : Anti-FTO antibody [EPR24440-12] (ab280081) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse heart tissue lysate
Lane 3 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 58 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Exposure time: 3 minutes.
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Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling FTO with ab280081 at 1/50 (11.98 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear and weak cytoplasmic staining in NIH/3T3 cell line is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling FTO with ab280081 at 1/500 dilution (0.1ug)/ (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab280081 has been referenced in 2 publications.
- Li S & Cao L Demethyltransferase FTO alpha-ketoglutarate dependent dioxygenase (FTO) regulates the proliferation, migration, invasion and tumor growth of prostate cancer by modulating the expression of melanocortin 4 receptor (MC4R). Bioengineered 13:5598-5612 (2022). PubMed: 34787056
- Li W et al. Comprehensive analysis of RNA m6A methylation in pressure overload-induced cardiac hypertrophy. BMC Genomics 23:576 (2022). PubMed: 35953789