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Recombinant SETDB1 protein

Expressed In

Baculovirus

Protein Species

Human

Cat No : 31452 31452

验证数据展示

  • Recombinant SETDB1 protein gel. SETDB1 protein was run on a 7.5% SDS-PAGE gel and stained with Coomassie Blue. MW: 145 kDa Purity: > 85%

    Recombinant SETDB1 protein gel. SETDB1 protein was run on a 7.5% SDS-PAGE gel and stained with Coomassie Blue. MW: 145 kDa Purity: > 85%

  • HTRF assay for Recombinant SETDB1 protein activity. 3 uM histone peptide H3 (1-21) was incubated with SETDB1 in reaction buffer including 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 uM SAM for 3 hours at room temperature. H3K9me2 antibody was used to detect reaction product.

    HTRF assay for Recombinant SETDB1 protein activity. 3 uM histone peptide H3 (1-21) was incubated with SETDB1 in reaction buffer including 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 uM SAM for 3 hours at room temperature. H3K9me2 antibody was used to detect reaction product.

  • HTRF assay for Recombinant SETDB1 protein activity. 3 uM histone peptide H3 (1-21) was incubated with SETDB1 in reaction buffer including 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 uM SAM for 3 hours at room temperature. H3K9me2 antibody was used to detect reaction product.

    HTRF assay for Recombinant SETDB1 protein activity. 3 uM histone peptide H3 (1-21) was incubated with SETDB1 in reaction buffer including 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 uM SAM for 3 hours at room temperature. H3K9me2 antibody was used to detect reaction product.

产品信息

Expressed In Baculovirus
Protein Species Human

Contents

Recombinant SETDB1 protein is supplied in 25 mM HEPES pH 7.5, 300 mM NaCl, 10% glycerol, 0.04% Triton X-100, 0.5 mM TCEP.

背景介绍

SETDB1 (SET Domain, Bifurcated 1) is a histone methyltransferase that methylates histone H3 at lysine 9. H3K9 methylation is associated with transcriptional repression and gene silencing through recruitment of HP1 proteins to methylated histones. H3K9 trimethylation is coordinated with DNA methylation, most likely through formation of a complex with MBD1 and ATF7IP that leads to transcriptional repression. SETDB1 activity is dependent on MBD1 and is heritably maintained through DNA replication by being recruited by CAF-1. SETDB1 is targeted to histone H3 by TRIM28, a factor recruited by KRAB zinc-finger proteins. Aberrant SETDB1 function is associated with the pathogenesis of Huntington Disease.

Application Notes

Recombinant SETDB1 is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling. HMT Assay Conditions: 3 uM H3 (1-21) peptide was incubated with different concentrations of recombinant SETDB1 protein in a 10 ul reaction system containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 uM SAM for 1 hour, then 10 ul H3K9me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

Protein Details

Recombinant SETDB1 protein was expressed in a baculovirus expression system as the full length protein (accession number NP_005245698.1) with an N-terminal FLAG-Tag. The molecular weight of the protein is 145 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.