Anti-FTO antibody [5-2H10] (ab92821)
![Western blot - Anti-FTO antibody [5-2H10] (ab92821)](https://www.abcam.com/ps/products/92/ab92821/Images/ab92821-280294-anti-fto-antibody-5-2h10-western-blot.jpg "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
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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
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Immunogen
Synthetic peptide from Human FTO.
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Positive control
- WB: Rat testes nuclear lysate. ICC/IF: Neuronal progenitor cells. Flow Cyt: SH-SY5Y cells.
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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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
Constituents: 0.238% HEPES, 0.01% BSA, 50% Glycerol, 0.87% Sodium chloride -
Concentration information loading... -
Purity
Protein G purified -
Purification notes
Protein G purified culture supernatant. -
Clonality
Monoclonal -
Clone number
5-2H10 -
Isotype
IgG2a -
Research areas
Associated products
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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 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).
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| ICC/IF | (1) |
1/100.
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| 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). |
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ICC/IF
1/100. |
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Flow Cyt
Use 1µg for 106 cells. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
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: 291905 Rat
- Omim: 610966 Human
- SwissProt: Q9C0B1 Human
- SwissProt: Q2A121 Rat
- Unigene: 528833 Human
- Unigene: 11858 Rat
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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 [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.
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![Flow Cytometry - Anti-FTO antibody [5-2H10] (ab92821)](https://www.abcam.com/ps/products/92/ab92821/Images/ab92821-108015-anti-fto-antibody-5-2h10-flow-cytometry.jpg)
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)](https://www.abcam.com/ps/products/92/ab92821/Images/ab92821-108016-anti-fto-antibody-5-2h10-immunofluorescence.jpg)
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)](https://www.abcam.com/ps/products/92/ab92821/Images/ab92821-108017-anti-fto-antibody-5-2h10-western-blot.jpg)
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
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (42)
ab92821 has been referenced in 42 publications.
- Wang A et al. m6A modifications regulate intestinal immunity and rotavirus infection. Elife 11:N/A (2022). PubMed: 35098923
- Zou L et al. N6-methyladenosine demethylase FTO suppressed prostate cancer progression by maintaining CLIC4 mRNA stability. Cell Death Discov 8:184 (2022). PubMed: 35397614
- Duan X et al. m6A demethylase FTO promotes tumor progression via regulation of lipid metabolism in esophageal cancer. Cell Biosci 12:60 (2022). PubMed: 35568876
- Li N et al. Targeting Ferroptosis as a Novel Approach to Alleviate Aortic Dissection. Int J Biol Sci 18:4118-4134 (2022). PubMed: 35844806
- Liu K et al. Exercise training ameliorates myocardial phenotypes in heart failure with preserved ejection fraction by changing N6-methyladenosine modification in mice model. Front Cell Dev Biol 10:954769 (2022). PubMed: 36120562