Recombinant JMJD2A / KDM4A protein

Aliases: Lysine Demethylase 4A, Jumonji C Domain-Containing Histone Demethylase 3A, JHDM3A
Catalog No: 31457 Format: 20 µg $410 Buy
Catalog No: 31857 Format: 1 mg $3,500 Buy
Expressed In: Baculovirus Protein Species: Human

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Contents

Full length recombinant JMJD2A / KDM4A protein was expressed in Sf9 cells and is supplied in 25 mM Hepes pH 7.5, 300 mM NaCl, 5% Glycerol, 0.04% Triton X-100, 0.2 mM TCEP. Please refer to product insert upon arrival for lot-specific concentration.

Background

KDM4A (lysine (K)-specific demethylase 4A), also known as JMJD2A (Jumonji Domain Containing 2A) is a nuclear protein that functions as a histone demethylase that preferentially demethylates di- and trimethylated lysine 9 (K9me3) and lysine 36 (K36me3) residues of histone H3, converting these trimethylated histone residues to mono- or dimethylated form. JMJD2A / KDM4A has no activity for monomethylated H3K9 and H3K36. KDM4A functions as a transcriptional repressor, participating in transcriptional repression of ASCL2 and E2F-responsive promoters via the recruitment of histone deacetylases and NCOR1, respectively. An additional KDM4A isoform, KDM4A Isoform 2, that lacks the N-terminal demethylase domain is crucial for muscle differentiation in promoting transcriptional activation of the MyoG gene by directing the removal of repressive chromatin marks at its promoter.

Application Notes

Recombinant JMJD2A / KDM4A 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 JMJD2A / KDM4A 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 JMJD2A / KDM4A (accession number NP_055478.2) was expressed in Sf9 cells and contains an N-terminal FLAG-Tag with an observed molecular weight of 125.3 kDa. The recombinant protein is >85% pure by SDS-PAGE.

References

This protein is described in the following references:
Zhang D et al. (2005) Mol Cell Biol. 25:6404-6414
Gray SG et al. (2005) J Biol Chem. 280:28507-28518
Cell Death Dis. (2018). 9(10): 1038. PMID: 30305606.

 
JMJD2A / KDM4A Coomassie gel

Recombinant JMJD2A / KDM4A protein gel.
JMJD2A / KDM4A protein was run on a 8% SDS-PAGE gel and stained with Coomassie blue.

JMJD2A / KDM4A activity assay

JMJD2A / KDM4A activity assay.
3.3 µM H3K9me3 peptide was incubated with JMJD2A / KDM4A in reaction buffer for 1 hour at room temperature. JMJD2A / KDM4A enzyme was used in a HTRF assay to determine enzyme linearity. Demethylated peptide (H3K9me2) was measured using H3K9me2-specific antibody.

JMJD2A / KDM4A activity assay

JMJD2A / KDM4A activity assay.
3.3 µM H3K9me3 peptide was incubated with 60 nM JMJD2A / KDM4A 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 negative control.

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.