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Recombinant EHMT2 (G9a) protein

Expressed In

Baculovirus

Protein Species

Human

Activ Motif logo

Cat No : 31410

Validation Data Gallery

  • Recombinant EHMT2 (G9a) protein gel. EHMT2 (G9a) protein was run on an 10% SDS-PAGE gel and stained with Coomassie Blue. MW: 134 kDa Purity: > 90%

    Recombinant EHMT2 (G9a) protein gel. EHMT2 (G9a) protein was run on an 10% SDS-PAGE gel and stained with Coomassie Blue. MW: 134 kDa Purity: > 90%

  • Recombinant EHMT2 (G9a) HTRF activity assay 1 μM histone H3 (1-21aa.) peptide was incubated with different concentrations of Recombinant EHMT2 (G9A) 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 3 hours, then 10 μl anti-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. Fluorenscence intensity at 620 nm and 665 nm were measured respectively. The final HTRF signal was obtained by formula: HTRF signal = F665/F620*10000.

    Recombinant EHMT2 (G9a) HTRF activity assay 1 μM histone H3 (1-21aa.) peptide was incubated with different concentrations of Recombinant EHMT2 (G9A) 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 3 hours, then 10 μl anti-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. Fluorenscence intensity at 620 nm and 665 nm were measured respectively. The final HTRF signal was obtained by formula: HTRF signal = F665/F620*10000.

  • Recombinant EHMT2 (G9a) HTRF activity assay 1 μM histone H3 (1-21aa.) peptide was incubated with different concentrations of Recombinant EHMT2 (G9A) 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 3 hours, then 10 μl anti-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. Fluorenscence intensity at 620 nm and 665 nm were measured respectively. The final HTRF signal was obtained by formula: HTRF signal = F665/F620*10000

    Recombinant EHMT2 (G9a) HTRF activity assay 1 μM histone H3 (1-21aa.) peptide was incubated with different concentrations of Recombinant EHMT2 (G9A) 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 3 hours, then 10 μl anti-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. Fluorenscence intensity at 620 nm and 665 nm were measured respectively. The final HTRF signal was obtained by formula: HTRF signal = F665/F620*10000

Product Information

Expressed In Baculovirus
Protein Species Human

Contents

Full length recombinant EHMT2 (G9a) protein expressed in Sf9 cells and supplied in 25 mM HEPES-NaOH pH 7.5, 300 mM NaCl, 10% glycerol, 0.04% Triton X-100, and 0.5 mM TCEP.

Background

EHMT2 (G9a) is a histone methyltransferase that specifically mono- and dimethylates Lys9 of histone H3 (H3K9me1 and H3K9me2, respectively) in euchromatin. H3K9me represents a specific tag for epigenetic transcriptional repression by initiating recruitment of HP1 proteins to methylated histones. EHMT2 (G9a) also mediates monomethylation of Lys56 of histone H3 (H3K56me1) in G1 phase and regulates DNA replication by promoting the interaction between histone H3 and PCNA. EHMT2 (G9a) also weakly methylates Lys27 of histone H3 (H3K27me). In addition, EHMT2 (G9a) is required for DNA methylation. However, the histone methyltransferase activity is not required for DNA methylation, suggesting that these 2 activities function independently. EHMT2 (G9a) is most likely targeted to histone H3 by different DNA-binding proteins like E2F6, MGA, MAX and/or DP1. EHMT2 (G9a) may also methylate histone H1. In addition to histone methyltransferase activity, EHMT2 (G9a) also methylates non-histone proteins. It has been shown to mediate dimethylation of Lys373 of p53 and also methylates CDYL, WIZ, ACIN1, DNMT1, HDAC1, ERCC6, KLF12 and itself.

Application Notes

Recombinant EHMT2 (G9A) protein is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

Protein Details

EHMT2 (G9a) is a histone methyltransferase that is specific to lysine 9 of histone H3 (H3K9). Methylation of H3K9 is a signal of transcriptional repression. Recombinant EHMT2 (G9a) protein (accession number NP_006700.3) was expressed in Sf9 cells and contains an N-terminal FLAG-Tag with a molecular weight of 134 kDa. The recombinant protein is > 90 % pure by SDS-PAGE.

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.