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Recombinant ALKBH5 protein

Expressed In

Baculovirus

Protein Species

Human

Cat No : 31589 31589

验证数据展示

  • Recombinant ALKBH5 protein gel Recombinant ALKBH5 was run on a 10% SDS-PAGE gel stained with Coomassie Blue. MW: 45.5 kDa Purity: > 90%

    Recombinant ALKBH5 protein gel Recombinant ALKBH5 was run on a 10% SDS-PAGE gel stained with Coomassie Blue. MW: 45.5 kDa Purity: > 90%

  • Dot blot for Recombinant ALKBH5 protein activity. 3 uM m6A ssDNA oligos (sequence: 5’-GTTGCCTGTTCGTGTTGG/m6A/CTTGCCTGT-3’) were incubated with different concentrations of ALKBH5 proteins in a 30 ul reaction system including 50 mM HEPES-NaOH pH 7.5, 100 uM 2-oxoglutarate, 100 uM ascorbate, 50 μM (NH4)2Fe(SO4)2·6H2O and 1 mM TCEP for 2 hours at room temperature. Then each reaction was concentrated to a final volume of 10 ul. 1 ul of each reaction was spotted onto a positively charged nylon membrane and products were detected by Dot blot assay with m6A antibody (Cat# 61495).

    Dot blot for Recombinant ALKBH5 protein activity. 3 uM m6A ssDNA oligos (sequence: 5’-GTTGCCTGTTCGTGTTGG/m6A/CTTGCCTGT-3’) were incubated with different concentrations of ALKBH5 proteins in a 30 ul reaction system including 50 mM HEPES-NaOH pH 7.5, 100 uM 2-oxoglutarate, 100 uM ascorbate, 50 μM (NH4)2Fe(SO4)2·6H2O and 1 mM TCEP for 2 hours at room temperature. Then each reaction was concentrated to a final volume of 10 ul. 1 ul of each reaction was spotted onto a positively charged nylon membrane and products were detected by Dot blot assay with m6A antibody (Cat# 61495).

产品信息

Expressed In Baculovirus
Protein Species Human

Contents

Recombinant ALKBH5 protein is supplied in 25 mM HEPES-NaOH pH 7.5, 300 mM NaCl, 10% glycerol, 0.04% Triton X-100, and 0.5 mM TCEP.

背景介绍

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.

Application Notes

Recombinant ALKBH5 protein is suitable for use in enzyme kinetics, inhibitor screening, and selectivity profiling. 3 uM m6A ssDNA oligos (DNA sequence: 5’-GTTGCCTGTTCGTGTTGG/m6A/ CTTGCCTGT-3’) were incubated with ALKBH5 protein in reaction buffer including 50 mM HEPES-NaOH pH 7.5, 100 uM 2-oxoglutarate, 100 uM ascorbate, 50 uM (NH4)2Fe(SO4)2·6H2O and 1 mM TCEP for 2 hours at room temperature. Dot-Blot with m6A antibody (Cat. No. 61495) was used for activity detection.

Protein Details

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.

Storage

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.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.