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Encompassing Amino Acids: METTL3: 2-580; METTL14: 2-456
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 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.
Description: 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.
Format: Aqueous buffer solution.
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 20% glycerol, 3mM DTT, 100ug/ml flag peptide
Genbank: METTL3: NM_019852; METTL14: NM_020961
Storage Stability: At least 6 months at -80°C.
Tags: METTL3: N-terminal FLAG-tag; METTL14: N-terminal His-tag
Uniprot: METTL3: Q86U44; METTL14: Q9HCE5
Warnings: Avoid freeze/thaw cycles. For optimal storage, aliquot target into smaller quantities after centrifugation and store at recommended temperature.
Biosafety Level: Not applicable (BSL-1)
References: 1. Liu J. et al. Nat. Chem. Biol. 2014. 10:93-952. Wang X. et al. Nature. 2016. 534:575-578
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