Human FTO, also known as Fat mass and obesity-associated protein, ALKBH9; BMIQ14; GDFD; KIAA1752, and alpha-ketoglutarate-dependent dioxygenase, GenBank Accession No. NM_001080432, a.a 32-505 (end), with N-terminal His-Tag and expressed in a Baculovirus infected Sf9 cell expression system. MW = 56 kDa.

Fat mass and obesity-associated protein, ALKBH9, BMIQ14, GDFD, KIAA1752, alpha-ketoglutarate-dependent dioxygenase, fto

0.0045 pmol/min/µg

Assay done using ELISA technology. 50 ul reaction mix containing HEPES-based buffer and 0-1500 ng FTO is added to the wells coated with the substrate. Incubate overnight at RT. Add antibody against N6-methyladenosine, incubate 1 hr. Then, add secondary HRP-labeled antibody and incubate 30 min. Finally, add HRP chemiluminsecent substrates and read luminescence.

40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 20% glycerol, 180 mM Imidazole

At least 6 months at –80°C

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

1. Frayling, T., et al., Science. 2007; 316(5826): 889-894.

2. Dina, C., et al., Nature Genetics. 2007; 39(6): 724-726.

Avoid freeze/thaw cycles. Protein may be diluted to ≥ 100 µg/ml in PBS + glycerol and remain stable for 6 months at -80°C.

Demethylase

Protein