Recombinant SETDB1 protein

Aliases: SET Domain Bifurcated 1, ERG-Associated Protein With SET Domain, Histone H3-K9 Methyltransferase 4
Catalog No: 31452 Format: 20 µg $505 Buy
Catalog No: 31852 Format: 1 mg $4,050 Buy

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Expressed In: Baculovirus Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

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 µM H3 (1-21) peptide was incubated with different concentrations of recombinant SETDB1 protein in a 10 µl reaction system containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 µM SAM for 1 hour, then 10 µl 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_001138887.1) with an N-terminal FLAG-Tag. The molecular weight of the protein is 145 kDa.

References

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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 µM 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 µM SAM for 3 hours at room temperature. H3K9me2 antibody was used to detect reaction product.

HTRF assay for Recombinant SETDB1 protein activity.
3 µM 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 µM SAM for 3 hours at room temperature. H3K9me2 antibody was used to detect reaction product.

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.