Recombinant Anti-FTO antibody [EPR6895] (ab124892)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6895] to FTO
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-FTO antibody [EPR6895]
See all FTO primary antibodies -
Description
Rabbit monoclonal [EPR6895] to FTO -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Synthetic peptide within Human FTO aa 450-550. The exact sequence is proprietary.
-
Positive control
- IHC-P: Human Breast, lung carcinoma and kidney tissues; WB: 293T, SH-SY5Y, Caco2, HEK-293 and BxPC3 lysates.
-
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
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 9.30 x 10 -13 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6895 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab124892 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) |
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
|
IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
Notes |
---|
WB
1/1000 - 1/10000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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. - 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
see all
Images
-
All lanes : Anti-FTO antibody [EPR6895] (ab124892) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : FTO knockout MCF7 cell lysate
Lane 3 : HEK-293 cell lysate
Lane 4 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 58 kDaFalse colour image of Western blot: Anti-FTO antibody [EPR6895] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab124892 was shown to bind specifically to FTO. A band was observed at 58 kDa in wild-type MCF7 cell lysates with no signal observed at this size in FTO knockout cell line. To generate this image, wild-type and FTO knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
-
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 - ab124892 observed at 58 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab124892 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. ab124892 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.
-
All lanes : Anti-FTO antibody [EPR6895] (ab124892) at 1/1000 dilution (Purified)
Lane 1 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 58 kDa
Observed band size: 58 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling FTO with purified ab124892 at 1:100 dilution (1.66 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast tissue sections labeling FTO with purified ab124892 at 1:100 dilution (1.66 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
-
ab124892, at 1/100 dilution, staining FTO in Paraffin-embedded Human Breast tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
-
All lanes : Anti-FTO antibody [EPR6895] (ab124892) at 1/1000 dilution
Lane 1 : 293T lysate
Lane 2 : SH-SY5Y lysate
Lane 3 : Caco2 lysate
Lane 4 : BxPC3 lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 58 kDa -
ab124892, at 1/100 dilution, staining FTO in Paraffin-embedded Human kidney tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (42)
ab124892 has been referenced in 42 publications.
- Udompatanakorn C et al. A study of RNA m6A demethylases in oral epithelial dysplasia and oral squamous cell carcinoma. J Oral Biol Craniofac Res 13:111-116 (2023). PubMed: 36582218
- Kim H et al. The m6A(m)-independent role of FTO in regulating WNT signaling pathways. Life Sci Alliance 5:N/A (2022). PubMed: 35169043
- Tsuchiya K et al. m6A demethylase ALKBH5 promotes tumor cell proliferation by destabilizing IGF2BPs target genes and worsens the prognosis of patients with non-small-cell lung cancer. Cancer Gene Ther 29:1355-1372 (2022). PubMed: 35318440
- Phillips S et al. Protocol for the generation of HIV-1 genomic RNA with altered levels of N6-methyladenosine. STAR Protoc 3:101616 (2022). PubMed: 35990737
- Garg R et al. Targeting FTO Suppresses Pancreatic Carcinogenesis via Regulating Stem Cell Maintenance and EMT Pathway. Cancers (Basel) 14:N/A (2022). PubMed: 36497402