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Recombinant KMT2D (MLL4) complex

商品コード: 31499 Format: 20 µg ¥77,000 Buy Now
商品コード: 31903 Format: 1 mg ¥648,000 Buy Now
Expressed In: E. coli Protein Species: Human

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for Recombinant KMT2D (MLL4) complexContents

Recombinant KMT2D (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. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant KMT2D (MLL4) complexBackground

KMT2D (MLL4) (Myeloid/lymphoid or mixed-lineage leukemia 4) is a Trithorax-group protein that function collectively to promote gene expression. KMT2D (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 β-globin locus transcription regulation by being recruited by Nfe2. KMT2D (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.

for Recombinant KMT2D (MLL4) complexApplication Notes

Recombinant KMT2D (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 KMT2D (MLL4) 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.

for Recombinant KMT2D (MLL4) complexProtein Details

Recombinant KMT2D (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 6×His-Tag and MW = 35 kDa; full length human ASH2L (GenBank Accession No. NM_001105214), N-terminal 6×His-Tag and MW = 61 kDa; full length human RbBP5 (GenBank Accession No. NM_005057), N-terminal 6×His-Tag and MW = 60 kDa, and full length human DPY30 (GenBank Accession No. NM_032574), N-terminal 6×His-Tag and MW =12 kDa, all individually expressed in E. coli cells. The recombinant protein is >85% pure by SDS-PAGE.

 
Recombinant KMT2D (MLL4) complex Coomassie gel

Recombinant KMT2D (MLL4) complex gel
MLL4 complex was run on an 10% SDS-PAGE gel and stained with Coomassie blue.

Recombinant KMT2D (MLL4) complex HTRF activity assay

Recombinant KMT2D (MLL4) complex HTRF activity assay
3.3 μM H3K4me0 (1-21aa) peptide was incubated with Recombinant KMT2D (MLL4) complex in reaction buffer for 3 hour at room temperature. Recombinant KMT2D (MLL4) complex was used in a HTRF assay to determine enzyme linearity. Methylated peptide (H3K4me2) was measured using H3K4me2-specific antibody.

Recombinant KMT2D (MLL4) complex MALDI-TOF activity assay

Recombinant KMT2D (MLL4) complex MALDI-TOF activity assay
3.3 uM H3K4me0 peptide was incubated with 20 nM Recombinant KMT2D (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

for Recombinant KMT2D (MLL4) complexStorage

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.

for Recombinant KMT2D (MLL4) complexGuarantee

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.

 

Technical Data Sheet

Recombinant KMT2D (MLL4) complex
 

Data Thumbnails

Recombinant KMT2D (MLL4) complex Coomassie gel

Recombinant KMT2D (MLL4) complex Coomassie gel.
(Click image to enlarge and see details).

Recombinant KMT2D (MLL4) complex HTRF activity assay

Recombinant KMT2D (MLL4) complex HTRF activity assay.
(Click image to enlarge and see details).

Recombinant KMT2D (MLL4) complex MALDI-TOF activity assay

Recombinant KMT2D (MLL4) complex MALDI-TOF activity assay.
(Click image to enlarge and see details).