Recombinant KMT2B (MLL4) complex

Expressed In

E. coli

Protein Species

Human

Cat No : 31499

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Validation Data Gallery

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  Recombinant KMT2B (MLL4) complex gel KMT2B (MLL4) complex was run on an 10% SDS-PAGE gel and stained with Coomassie blue.

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  Recombinant KMT2B (MLL4) complex HTRF activity assay 3.3 uM H3K4me0 (1-21aa) peptide was incubated with Recombinant KMT2B (MLL4) complex in reaction buffer for 3 hour at room temperature. Recombinant KMT2B (MLL4) complex was used in a HTRF assay to determine enzyme linearity. Methylated peptide (H3K4me2) was measured using H3K4me2-specific antibody.

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  Recombinant KMT2B (MLL4) complex MALDI-TOF activity assay 3.3 uM H3K4me0 peptide was incubated with 20 nM Recombinant KMT2B (MLL4) complex in reaction buffer for 3 hours at room temperature. The reaction product was detected by MALDI-TOF. Single 3.3 uM H3K4me0 peptide was used as negative control. Catalytic Ability: 50 turnovers/ enzyme molecule

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Product Information

• Expressed In: E. coli
• Protein Species: Human

Contents

Recombinant KMT2B (MLL4) complex was expressed in E.coli cells and is supplied in 25 mM Tris pH 8.0, 300 mM NaCl, 5% Glycerol, 0.04% Triton X-100.

Background

KMT2B (MLL4) (Myeloid/lymphoid or mixed-lineage leukemia 4) is a Trithorax-group protein that function collectively to promote gene expression. KMT2B (MLL4) is a histone methyltransferase that methylates the Lysine 4 position of histone H3. In particular, the SET domain is a conserved C-terminal domain that characterizes proteins of the MLL (mixed-lineage leukemia) family. The SET domain is responsible for its histone methyltransferase activity which mediates chromatin modifications associated with epigenetic transcriptional activation. H3K4 methylation represents a specific epigenetic tag for transcriptional activation. MLL4 plays a central role in b-globin locus transcription regulation by being recruited by Nfe2. KMT2B (MLL4) is also required during the transcriptionally active period of oocyte growth for the establishment and/or maintenance of bulk H3K4 trimethylation (H3K4me3), global transcriptional silencing that precedes resumption of meiosis, oocyte survival and normal zygotic genome activation. MLL4 may also be involved in human cancer as its expression has been shown to be upregulated in solid tumor cell lines.

Application Notes

Recombinant KMT2B (MLL4) complex is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling. HMT Assay Conditions: 3.3 μM H3K4me0 (1-21aa) peptide was incubated with different concentrations of recombinant KMT2B (MLL4) complex in reaction buffer containing 50 mM TrisCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 100 uM SAM for 3 hours at room temperature. Activity was detected by HTRF and MALDI-TOF.

Protein Details

Recombinant KMT2B (MLL4) complex contains amino acids 2551 - 2715 of the human MLL4 protein (accession number NP_055542.1) with N-terminal GST-Tag and MW = 47.6 kDa; full length human WDR5 (GenBank Accession No.NM_017588), with N-terminal 6xHis Tag and MW = 35 kDa; full length human ASH2L (GenBank Accession No. NM_001105214), N-terminal 6xHis Tag and MW = 61 kDa; full length human RbBP5 (GenBank Accession No. NM_005057), N-terminal 6xHis Tag and MW = 60 kDa, and full length human DPY30 (GenBank Accession No. NM_032574), N-terminal 6xHis Tag and MW =12 kDa, all individually expressed in E. coli cells. The recombinant protein is >85% 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.