Recombinant SETD6 Enzyme Description: Recombinant full-length human SETD6 protein (SET6, accession Q8TBK2, amino acids 1-473), containing an N-terminal GST tag, expressed in E. coli. SETD6 is a SET-domain containing protein lysine methyltransferase, catalyzing the mono-methylation of RelA (p65, NFkB3) at lysine 310. Methylation of RelA attenuates RelA-dependant transcriptional pathways, including inflammatory responses in primary immune cells. SETD6 is downregulated in a diseases of hyper inflammation.
Recombinant SETD6 Enzyme Formulation: Recombinant GST-SETD6 (2.2 µg/µl) 100 mM Tris pH 8.0, 10mM 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 SETD6, human is useful for RelA/p65 methylation experiments, enzyme kinetics and inhibitor screening. Use of 2-5 µg SETD6 per reaction with 1 µg recombinant RelA as a substrate is recommended.
References: Levy D et al (2011). Nat Immunol 12: 29-36. Link
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Enzyme Activity Data: Recombinant SETD6, human (4 µg) was used in a methylation assay with 1 µg recombinant RelA and radioactive SAM and the reaction was run on a PAGE gel. Lane 1: GST control. Lane 2: GST-SETD6. Top panel: Autoradiograh. Bottom Panel: RelA immunoblot. (Click image to enlarge)
Protein Gel Data: Recombinant SETD6, 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)