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Human FTO ELISA Kit is a single-wash 90-min Simplestep used to quantify Human FTO with a sensitivity of 11.96 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

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Images

Sandwich ELISA - Human FTO ELISA Kit (AB309316), expandable thumbnail
  • Sandwich ELISA - Human FTO ELISA Kit (AB309316), expandable thumbnail

Key facts

Sample types
Cell Lysate, Tissue Extracts
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
54.688 - 3500 pg/mL
Assay time
1h 30m
Sensitivity
= 11.96 pg/mL

Reactivity data

Application
sELISA
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

RNA demethylase that mediates oxidative demethylation of different RNA species, such as mRNAs, tRNAs and snRNAs, and acts as a regulator of fat mass, adipogenesis and energy homeostasis (PubMed:22002720, PubMed:25452335, PubMed:26457839, PubMed:26458103, PubMed:28002401, PubMed:30197295). Specifically demethylates N(6)-methyladenosine (m6A) RNA, the most prevalent internal modification of messenger RNA (mRNA) in higher eukaryotes (PubMed:22002720, PubMed:25452335, PubMed:26457839, PubMed:26458103, PubMed:30197295). M6A demethylation by FTO affects mRNA expression and stability (PubMed:30197295). Also able to demethylate m6A in U6 small nuclear RNA (snRNA) (PubMed:30197295). Mediates demethylation of N(6),2'-O-dimethyladenosine cap (m6A(m)), by demethylating the N(6)-methyladenosine at the second transcribed position of mRNAs and U6 snRNA (PubMed:28002401, PubMed:30197295). Demethylation of m6A(m) in the 5'-cap by FTO affects mRNA stability by promoting susceptibility to decapping (PubMed:28002401). Also acts as a tRNA demethylase by removing N(1)-methyladenine from various tRNAs (PubMed:30197295). Has no activity towards 1-methylguanine (PubMed:20376003). Has no detectable activity towards double-stranded DNA (PubMed:20376003). Also able to repair alkylated DNA and RNA by oxidative demethylation: demethylates single-stranded RNA containing 3-methyluracil, single-stranded DNA containing 3-methylthymine and has low demethylase activity towards single-stranded DNA containing 1-methyladenine or 3-methylcytosine (PubMed:18775698, PubMed:20376003). Ability to repair alkylated DNA and RNA is however unsure in vivo (PubMed:18775698, PubMed:20376003). Involved in the regulation of fat mass, adipogenesis and body weight, thereby contributing to the regulation of body size and body fat accumulation (PubMed:18775698, PubMed:20376003). Involved in the regulation of thermogenesis and the control of adipocyte differentiation into brown or white fat cells (PubMed:26287746). Regulates activity of the dopaminergic midbrain circuitry via its ability to demethylate m6A in mRNAs (By similarity). Plays an oncogenic role in a number of acute myeloid leukemias by enhancing leukemic oncogene-mediated cell transformation: acts by mediating m6A demethylation of target transcripts such as MYC, CEBPA, ASB2 and RARA, leading to promote their expression (PubMed:28017614, PubMed:29249359).

Alternative names

What's included?

1 x 96 Tests
Components
10X Human FTO Capture Antibody
1 x 600 µL
10X Human FTO Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
5X Cell Extraction Buffer PTR (ab193970)
1 x 10 mL
Antibody Diluent CPR2
1 x 6 mL
Human FTO Lyophilized Recombinant Protein
1 x 2 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
1 x 12 mL
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 1 Unit
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

Recommended products

Human FTO ELISA Kit is a single-wash 90-min Simplestep used to quantify Human FTO with a sensitivity of 11.96 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

Key facts

Sample types
Cell Lysate, Tissue Extracts
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
54.688 - 3500 pg/mL
Assay time
1h 30m
Assay Platform
Pre-coated microplate (12 x 8 well strips)
Sensitivity
= 11.96 pg/mL
General recovery
108%

Precision

Intra assay

Sample
Extract
n
8
C.V.
7.9

Inter assay

Sample
Extract
n
3
C.V.
3.6

Recovery

Sample specific recovery

Sample type
Cell Lysate
Average %
= 108
Range
100 - 113 %
Sample type
Tissue Extracts
Average %
= 112
Range
104 - 122 %

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Notes

Human FTO SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of FTO protein in human serum, plasma, saliva and cell culture supernatant. Quantitate Human FTO with 11.96 pg/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The FTO protein also referred to as Alpha-Ketoglutarate-Dependent Dioxygenase or Fat Mass and Obesity-Associated protein functions as an enzyme involved in demethylating nucleic acids specifically N6-methyladenosine (m6A) in RNA. This process impacts RNA stability and translation. FTO has a molecular weight of approximately 58 kDa. Higher expression levels of FTO are found in the brain including the hypothalamus which is an area known for regulating energy balance and appetite.

Biological function summary

FTO influences energy homeostasis and adipogenesis. It acts as an important player in metabolic regulation and gene expression by modulating the m6A RNA modification. While FTO operates largely on its own its demethylation activity may interact with other RNA binding proteins that recognize m6A-modified RNAs integrating it into larger regulatory frameworks for RNA metabolism and cellular responses.

Pathways

FTO is an important component in the regulation of energy balance and metabolism. The protein is closely associated with the mTOR signaling pathway which is essential for controlling cell growth proliferation and survival in response to nutrient availability. FTO also interacts with the leptin signaling pathway relating to its role in appetite regulation and energy expenditure.

Associated diseases and disorders

FTO has significant connections to obesity and Type 2 diabetes. Genetic variants within the FTO gene correlate with risk for these conditions linking its enzymatic activity to alterations in energy balance and insulin sensitivity. FTO's role in Type 2 diabetes also highlights its interaction with insulin signaling proteins demonstrating its broader influence on glucose metabolism.

Product promise

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In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

2 product images

  • Sandwich ELISA - Human FTO ELISA Kit (ab309316), expandable thumbnail

    Sandwich ELISA - Human FTO ELISA Kit (ab309316)

    Interpolated concentration of native FTO was measured in duplicate at different sample concentrations. Undiluted samples are as follows: HepG2 cell extract 25 µg/mL, NIH/3T3 cell extract 125 µg/mL, and human thyroid tissue extract 250 µg/mL.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR.

  • Sandwich ELISA - Human FTO ELISA Kit (ab309316), expandable thumbnail

    Sandwich ELISA - Human FTO ELISA Kit (ab309316)

    Example of human FTO standard curve. Background-subtracted data values (mean +/- SD) are graphed.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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