Recombinant DNMT3A / DNMT3L complex

Aliases: DNA Methyltransferase 3A / DNA Methyltransferase 3-like protein
Catalog No: 31415 Format: 20 µg $505 Buy
Catalog No: 31815 Format: 1 mg $4,300 Buy

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Expressed In: Baculovirus 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

DNA methyltransferase proteins such as DNMT3A are involved in DNA methylation, in which a methyl group is added to a cytosine residue on DNA, commonly at the C5 position of a CpG dinucleotide. DNMT3A has been shown to be important in the regulation of specific patterns of DNA methylation. Methylation of mammalian DNA has long been recognized to play a major role in a number of cellular functions such as embryonic development, genetic imprinting, X chromosome inactivation and control of gene expression. DNA methylation is generally associated with transcriptional repression.

Three families of DNMTs have been identified: DNMT1, DNMT2 and DNMT3. The DNMT3 family, containing two active methyltransferases, DNMT3A & DNMT3B, and one DNMT3-Like protein (DNMT3L), establishes the initial CpG methylation pattern de novo and shows the same propensity for methylating unmethylated duplex DNA as for hemi-methylated DNA. DNMT3A and DNMT3B are encoded by two distinct genes and are strongly expressed in embryonic stem cells. DNMT3A is ubiquitous in mammalian cells. DNMT3L is a nuclear protein with similarity to the DNMT family of proteins, but does not function as a DNA methyltransferase. It serves as an essential regulator of DNMT3A and DNMT3B, recruiting them to sites for deposition of de novo DNA methylation.

Application Notes

Recombinant DNMT3A / DNMT3L complex is suitable for use in enzyme kinetics, inhibitor screening, and selectivity profiling.

DNMT Activity Assay Conditions: Assay was performed using DNMT Activity / Inhibition Assay, Active Motif Catalog No. 55006, for 1 hour at 37°C.

Protein Details

Recombinant DNMT3A / DNMT3L complex that includes human DNMT3A amino acids 214-912 (accession number NP_783328.1) with an N-terminal 6xHis Tag and FLAG-Tag and full length DNMT3L (accession number NP_787063.1) with an N-terminal 6xHis Tag was expressed in Sf9 cells. The molecular weights of DNMT3A and DNMT3L are 83.8 kDa and 46.6 kDa, respectively. The recombinant protein is >95% pure by SDS-PAGE.

 

Recombinant DNMT3A / DNMT3L complex gel
Recombinant DNMT3A / DNMT3L complex was run on an 8% SDS-PAGE gel and stained with Coomassie blue.

Recombinant DNMT3A / DNMT3L complex activity assay
Recombinant DNMT3A / DNMT3L complex activity was measured using DNMT Activity / Inhibition Assay, Active Motif Catalog No. 55006, for 1 hour at 37°C

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.