Recombinant SETD7 protein

Aliases: KMT7, SET7, SET9, SET7/9
Catalog No: 31496 Format: 20 µg $410 Buy
Expressed In: Baculovirus Protein Species: Human

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Contents

Full length recombinant SETD7 protein was expressed in Sf9 cells and is supplied in 25 mM Hepes pH 7.5, 300 mM NaCl, 5% Glycerol, 0.04% Triton X-100, 0.2 mM TCEP. Please refer to product insert upon arrival for lot-specific concentration.

Background

SETD7 (SET Domain Containing (Lysine Methyltransferase) 7) is a histone methyltransferase that specifically monomethylates ‘Lys-4’ of histone H3. H3 ‘Lys-4’ methylation represents a specific tag for epigenetic transcriptional activation. Plays a central role in the transcriptional activation of genes such as collagenase or insulin. Recruited by IPF1/PDX-1 to the insulin promoter, leading to activate transcription. Has also methyltransferase activity toward non-histone proteins such as p53/TP53, TAF10, and possibly TAF7 by recognizing and binding the [KR]-[STA]-K in substrate proteins. Monomethylates ‘Lys-189’ of TAF10, leading to increase the affinity of TAF10 for RNA polymerase II. Monomethylates ‘Lys-372’ of p53/TP53, stabilizing p53/TP53 and increasing p53/TP53-mediated transcriptional activation.

Application Notes

Recombinant SETD7 is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

HMT Assay Conditions: 1 μM H3K4me0 (1-21aa) peptide was incubated with different concentrations of recombinant SETD7 protein in reaction buffer containing 50 mM TrisCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 100 μM SAM for 2 hours at room temperature. Activity was detected by HTRF and Fluorography.

Protein Details

Recombinant SETD7 protein (accession number NP_085151.1) was expressed in Sf9 cells and contains an N-terminal FLAG-Tag with a molecular weight of 41.7 kDa. The recombinant protein is >80% pure by SDS-PAGE.

 
SETD7 protein Coomassie gel

Recombinant SETD7 protein gel.
SETD7 protein was run on an 8% SDS-PAGE gel and stained with Coomassie blue.

SETD7 protein HTRF activity assay

Recombinant SETD7 protein HTRF activity assay.
1 μM H3K4me0 (1-21aa) peptide was incubated with SETD7 in reaction buffer for 2 hour at room temperature. SETD7 enzyme was used in a HTRF assay to determine enzyme linearity. Methylated peptide (H3K4me1) was measured using H3K4me1-specific antibody.

SETD7 protein activity assay

Recombinant SETD7 protein activity assay.
2 ug Calf Thymus Octamers was incubated with 0.5 ug SETD7 protein in reaction buffer containing radioactive 3H-SAM for 2 hour at room temperature. Activity was detected by fluorography.

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.