Recombinant DOT1L Catalytic Domain Enzyme Description: Recombinant human DOT1 protein, catalytic domain (DOT1L, KMT4, accession Q8TEK3, amino acids 1-472), containing an N-terminal GST tag, expressed in E. coli. DOT1 is a histone methyltransferase, catalyzing the methylation of histone H3 at lysine 79 on nucleosomal substrates.
Recombinant DOT1L Catalytic Domain Enzyme Formulation: Recombinant GST-DOT1L (1 µg/µl) 100 mM Tris pH 8.0, 5 mM glutathione and 25% glycerol.
Storage and Stability: Stable for six months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.
Application Notes: Recombinant DOT1L Catalytic Domain, human is useful for histone H3 methylation experiments, enzyme kinetics and inhibitor screening. Use of 3-5 µg DOT1L per reaction with HeLa or recombinant nucleosomes as a substrate is recommended.
References: Kuo AJ et al (2011). Mol Cell 44: 609-620. Link
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Enzyme Activity Data: Recombinant DOT1L Catalytic Domain, human (5 µg) was used in a methylation assay with 1 µg recombinant Xenopus nucleosomes and radioactive SAM and the reaction was run on a PAGE gel. Lane 1: GST only vector. Lane 2: GST-DOT1L vector. Left Panel: Coomassie stained gel. Right panel: Autoradiograph. (Click image to enlarge)
Protein Gel Data: Recombinant DOT1L Catalytic Domain, human (1 µg) run on a PAGE gel and stained with Coomassie blue. Migration and molecular weight of protein standards is indicated. (Click image to enlarge)