Key features and details
- Mouse monoclonal [5-2H10] to FTO
- Suitable for: WB, ICC/IF, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG2a
Product nameAnti-FTO antibody [5-2H10]
See all FTO primary antibodies
DescriptionMouse monoclonal [5-2H10] to FTO
Tested applicationsSuitable for: WB, ICC/IF, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Sheep, Horse, Cow, Dog, Non human primates
Synthetic peptide from Human FTO.
- WB: Rat testes nuclear lysate. ICC/IF: Neuronal progenitor cells. Flow Cyt: SH-SY5Y cells.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferConstituents: 0.238% HEPES, 0.01% BSA, 50% Glycerol, 0.87% Sodium chloride
Concentration information loading...
PurityProtein G purified
Purification notesProtein G purified culture supernatant.
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipoprotein metabolism
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.
|WB||1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).|
|Flow Cyt||Use 1µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
FunctionDioxygenase 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 specificityUbiquitously expressed, with relatively high expression in adrenal glands and brain; especially in hypothalamus and pituitary.
Involvement in diseaseDefects 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 similaritiesBelongs to the fto family.
DomainThe 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.
- Information by UniProt
- AlkB homolog 9 antibody
- ALKBH9 antibody
- Alpha-ketoglutarate-dependent dioxygenase FTO antibody
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.
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.
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.
Anti-FTO antibody [5-2H10] (ab92821) at 1/1000 dilution + Rat testes lysate
Predicted band size: 58 kDa
Observed band size: 58 kDa
ab92821 has been referenced in 13 publications.
- Tang X et al. The role of the fat mass and obesity-associated protein in the proliferation of pancreatic cancer cells. Oncol Lett 17:2473-2478 (2019). PubMed: 30719115
- Song H et al. METTL3 and ALKBH5 oppositely regulate m6A modification of TFEB mRNA, which dictates the fate of hypoxia/reoxygenation-treated cardiomyocytes. Autophagy 15:1419-1437 (2019). PubMed: 30870073
- Huang B et al. Cyclophosphamide Regulates N6-Methyladenosine and m6A RNA Enzyme Levels in Human Granulosa Cells and in Ovaries of a Premature Ovarian Aging Mouse Model. Front Endocrinol (Lausanne) 10:415 (2019). PubMed: 31316467
- Li J et al. Elevated N-methyltransferase expression induced by hepatic stellate cells contributes to the metastasis of hepatocellular carcinoma via regulation of the CD44v3 isoform. Mol Oncol 13:1993-2009 (2019). PubMed: 31294922
- Tan B et al. Viral and cellular N6-methyladenosine and N6,2'-O-dimethyladenosine epitranscriptomes in the KSHV life cycle. Nat Microbiol 3:108-120 (2018). PubMed: 29109479