A representative Technical Data Sheet (TDS) is provided here. Please refer to your lot-specific TDS upon arrival for specific buffer contents and lot-specific protein concentration.
N6-methylated adenine (m6A) is prevalent in nearly all RNA types and can be found in all organisms from bacteria to humans. It preferentially appears around stop codons and within long internal exons in mammalian messager RNAs. m6A plays an important role in the efficiency of mRNA splicing, processing, translation efficiency, editing and mRNA stability. m6A also appears in other RNA molecules, such as primary miRNA (pri-miRNAs).
ALKBH5 (AlkB Homolog 5, RNA Demethylase, also known as OFOXD1, ABH5) specifically demethylates m6A RNA. ALKBH5 significantly affects mRNA export and RNA metabolism as well as the assembly of mRNA processing factors in nuclear speckles via demethylation activity.
Recombinant ALKBH5 protein is suitable for use in enzyme kinetics, inhibitor screening, and selectivity profiling.
3 µM m6A ssDNA oligos (DNA sequence: 5’-GTTGCCTGTTCGTGTTGG/m6A/ CTTGCCTGT-3’) were incubated with ALKBH5 protein in reaction buffer including 50 mM HEPES pH 7.5, 100 µM 2OG, 100 µM Ascorbate, 50 µM (NH4)2Fe(SO4)2·6H2O and 1 mM TCEP for 2 hours at room temperature. Dot-Blot was used for activity detection.
Recombinant ALKBH5 protein was expressed in a baculovirus expression system as the full length protein (accession number NP_060228.3) with an N-terminal FLAG-Tag. The molecular weight of ALKBH5 is 45.5 kDa.
Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.