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Encompassing Amino Acids: 1129-2002
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Add 50 µl reaction mix containing a-ketoglutarate, TET enzyme, ATP, and TET SA assay buffer (BPS) to wells coated with the substrate. TET SA buffer contains saturable concentrations of iron, a-KG, and ATP. Incubate 3hrs at 37°C. Add antibodyagainst hydroxymethylated cytosine, incubate 1 hr. Then add secondary HRP labeled antibody and incubate 30 min. Finally, add HRP chemiluminescent substrates and read luminescence.
Description: Human Ten-eleven translocation 2 protein (TET2), methylcytosine dioxygenase TET2, GenBank Accession # NM_001127208, a.a. 1129-2002 (end) with N-terminal FLAG-tag, C-terminal His-tag. MW = 100 kDa, expressed in Sf9 cells via a baculovirus expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 0.04% Tween-20, 20% glycerol, and 3 mM DTT.
Genbank: NM_001127208
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C. Avoid freeze/thaw cycles.
Tags: N-terminal FLAG-tag, C terminal His tag
Uniprot: Q6N021
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1.Myunggon, K., et al., Nature. 2013;497: 122-126.2.Shinsuke, I, et al., Nature. 2010; 466: 1129-1133.