Recombinant JMJD2D / KDM4D protein

Aliases: Lysine Demethylase 4D, Jumonji Domain-Containing Protein 2D, JHDM3D
Catalog No: 31459 Format: 20 µg $485 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

KDM4D (lysine (K)-specific demethylase 4D), also known as JMJD2D (Jumonji Domain Containing 2D) is a protein that functions as a histone demethylase that preferentially demethylates di- and trimethylated lysine 9 residues of histone H3, while it has no activity on monomethylated H3K9 residues.

Application Notes

Recombinant JMJD2D / KDM4D is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

Histone Demethylase Assay Conditions: 50 mM HEPES pH 7.5, 0.02% Triton X100, 100 μM 2OG, 100 μM Ascorbate, 50 μM (NH4)2Fe(SO4)2·6H2O, 1 mM TCEP, 100 nM Recombinant JMJD2D / KDM4D protein, and 3.3 μM H3K9me3 (aa 1-21) peptide at 2 hours at room temperature. MALDI-TOF was used for detection.

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Protein Details

Recombinant JMJD2D / KDM4D (accession number NP_060509.2) was expressed in Sf9 cells and contains an N-terminal FLAG-Tag with an observed molecular weight of 63.2 kDa. The recombinant protein is >60% pure by SDS-PAGE.

References

This protein is described in the following references:
Fodor BD et al. (2006) Genes Dev. 20:1557-1562
Beyer S et al. (2008) J Biol Chem. 283:36542-36552. doi: 10.1074/jbc.M804578200

 

Recombinant JMJD2D / KDM4D protein gel.
JMJD2D / KDM4D protein was run on an SDS-PAGE gel and stained with Coomassie Blue.

MALDI-TOF Assay for JMJD2D / KDM4D activity .
3.3 µM H3K9me3 peptide was incubated with 100 nM JMJD2D / KDM4D in reaction buffer for 2 hours at room temperature. The reaction product was detected by MALDI-TOF. Single 3.3 µM H3K9me3 peptide was used as a negative control.
Catalytic Ability: >66 turnovers / enzyme molecule.

Recombinant JMJD2D / KDM4D activity using AlphaLISA.
JMJD2D / KDM4D was used in an AlphaLISA assay to determine enzyme titration. An IC50 dose response assessment of reference compounds 2,4-PDCA and N-OG is shown. This data was generated and kindly provided courtesy of ChemPartner.

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.