Recombinant KMT2D (MLL2) complex

Catalog No: 31498 Format: 20 µg $475 Buy
Catalog No: 31902 Format: 1 mg $3,200 Buy

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Expressed In: E. coli 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

KMT2D (MLL2) complex (Myeloid/lymphoid or mixed-lineage leukemia 2) is a Trithorax-group protein that function collectively to promote gene expression. KMT2D (MLL2) 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. MLL2 is part of a large protein complex called ASCOM, which has been shown to be a transcriptional regulator of the β-globin and estrogen receptor genes. Mutations in this gene have been shown to be a cause of Kabuki syndrome.

Application Notes

Recombinant KMT2D (MLL2) 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 KMT2D (MLL2) complex 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 3 hours at room temperature. Activity was detected by HTRF and MALDI-TOF.

Protein Details

Recombinant KMT2D (MLL2) complex contains amino acids 5355 - 5537 of the human MLL2 protein (accession number NP_003473.3) 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 >80% pure by SDS-PAGE.

 

Recombinant KMT2D (MLL2) complex gel
Recombinant KMT2D (MLL2) complex was run on an 10% SDS-PAGE gel and stained with Coomassie blue.

Recombinant KMT2D (MLL2) complex HTRF activity assay
3.3 µM H3K4me0 (1-21aa) peptide was incubated with KMT2D (MLL2) complex in reaction buffer for 3 hour at room temperature. KMT2D (MLL2) complex was used in a HTRF assay to determine enzyme linearity. Methylated peptide (H3K4me2) was measured using H3K4me2-specific antibody.

Recombinant KMT2D (MLL2) complex MALDI-TOF activity assay
3.3 µM H3K4me0 peptide was incubated with 10 nM KMT2D (MLL2) 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: ~450 turnovers/ enzyme molecule

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.