Recombinant ALKBH2 protein

Catalog No: 81129 Format: 100 µg ¥62,000 Buy
Catalog No: 81829 Format: 1 mg ¥400,000 Buy
Expressed In: E. coli Protein Species: Human

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Contents

Recombinant ALKBH2 protein is supplied in 25 mM Tris pH 8.0, 300 mM NaCl, 20% glycerol, 0.5 mM TCEP.

Background

ALKBH2 (AlkB Homolog 2, Alpha-Ketoglutarate Dependent Dioxygenase) is a member of ALKBH protein family. It can repair alkylated DNA and RNA containing 1-methyladenine and 3-methylcytosine by oxidative demethylation. ALKBH2 can also repair alkylated DNA containing 1-ethenoadenine (in vitro). Has strong preference for double-stranded DNA. It can release modified bases from both DNA and RNA. The oxidative demethylation requires molecular oxygen, alpha-ketoglutarate and iron.

Application Notes

Recombinant ALKBH2 protein is suitable for use in enzyme kinetics, inhibitor screening, and selectivity profiling.

Protein Details

Full length ALKBH2 protein (accession number NP_001001655.1) was expressed in E. coli cells with an N-terminal 6xHis-Tag. The molecular weight of ALKBH2 is 33.1 kDa.

 

Recombinant ALKBH2 protein
10% SDS-PAGE Coomassie staining
MW: 33.1 kDa
Purity: >82%

ALKBH2 dioxygenase activity assay
1 µg of ssDNA oligos (5’-AAAGCAG(1mA)ATTCGAAAAAGCGAAA-3’) was incubated with 0 µg, 0.5 µg, 1 µg, 2 µg, 4 µg of ALKBH2 in 50 µl reaction system including 50 mM HEPES-NaOH pH 8.0, 50 μM Fe(NH4)2(SO4)2, 1 mM 2-oxoglutarate, 2 mM ascorbate and 1 mM TCEP for 30 min at 37°C. Then ssDNA oligos were annealed with equimolar of non-methylated complement strand followed by 1 µg EcoRI digestion for 45 min at 37°C. 1/4 reaction products were run on a 20% Native PAGE gel and stained by ethidium bromide.

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 for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.