Complex of Human Methyltransferase-like 3 (METTL3), GenBank Accession No. NM_019852, a.a. 2-580 (end) with N-terminal FLAG-tag, MW = 66 kDa and Methyltransferase-like 14 (METTL14), Genbank Accession No. NM_020961 a.a. 2-456 (end), with N-terminal His-tag, MW = 53 kDa, coexpressed by baculovirus in Sf9 insect cells.

Human Methyltransferase-like 3, METTL3; Methyltransferase-like 14, RNA

Add 50μl reaction mix containing S-adenosylmethionine, METTL enzyme, and METTL buffer to wells coated with the substrate. Incubate overnight at room temperature. Add antibody against methylated adenosine, incubate 1 hr. Then, add secondary HRP-labeled antibody and incubate 30 min. Finally, add HRP chemiluminescent substrates and read luminescence.

40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 20% glycerol, 3mM DTT, 100ug/ml flag peptide

Aqueous buffer solution.

At least 6 months at –80°C.

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

1. Liu J. et al. Nat. Chem. Biol. 2014. 10:93–95

2. Wang X. et al. Nature. 2016.  534:575-578 

Avoid freeze/thaw cycles. For optimal storage, aliquot target into smaller quantities after centrifugation and store at recommended temperature.

Methyltransferase

Protein