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Recombinant JMJD2A / KDM4A protein

Expressed In

Baculovirus

Protein Species

Human

Activ Motif logo

Cat No : 31457

Validation Data Gallery

  • Recombinant JMJD2A / KDM4A protein gel. JMJD2A / KDM4A protein was run on a 10% SDS-PAGE gel and stained with Coomassie blue. MW: 125.3 kDa Purity: > 85%

    Recombinant JMJD2A / KDM4A protein gel. JMJD2A / KDM4A protein was run on a 10% SDS-PAGE gel and stained with Coomassie blue. MW: 125.3 kDa Purity: > 85%

  • HTRF assay for JMJD2A / KDM4A activity 1 uM H3K9me3 peptide was incubated with different concentrations of JMJD2A / KDM4A protein in a 10 ul reaction system containing 50 mM HEPES-NaOH pH 7.5, 1 mM TCEP, 50 uM 2-OG, 50 uM Ascorbate and 25 uM (NH4)2Fe(SO4)2·6H2O for 1 hr, then 10 ul H3K9me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

    HTRF assay for JMJD2A / KDM4A activity 1 uM H3K9me3 peptide was incubated with different concentrations of JMJD2A / KDM4A protein in a 10 ul reaction system containing 50 mM HEPES-NaOH pH 7.5, 1 mM TCEP, 50 uM 2-OG, 50 uM Ascorbate and 25 uM (NH4)2Fe(SO4)2·6H2O for 1 hr, then 10 ul H3K9me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

  • HTRF assay for JMJD2A / KDM4A activity 1 uM H3K9me3 peptide was incubated with different concentrations of JMJD2A / KDM4A protein in a 10 ul reaction system containing 50 mM HEPES-NaOH pH 7.5, 1 mM TCEP, 50 uM 2-OG, 50 uM Ascorbate and 25 uM (NH4)2Fe(SO4)2·6H2O for 1 hr, then 10 ul H3K9me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

    HTRF assay for JMJD2A / KDM4A activity 1 uM H3K9me3 peptide was incubated with different concentrations of JMJD2A / KDM4A protein in a 10 ul reaction system containing 50 mM HEPES-NaOH pH 7.5, 1 mM TCEP, 50 uM 2-OG, 50 uM Ascorbate and 25 uM (NH4)2Fe(SO4)2·6H2O for 1 hr, then 10 ul H3K9me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

Product Information

Expressed In Baculovirus
Protein Species Human

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, 10% Glycerol, 0.04% Triton X-100, 0.5 mM TCEP.

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

This product was manufactured as described in Protein Details. Where possible, Active Motif has developed functional or activity assays for recombinant proteins. Additional characterization such as enzyme kinetic activity assays, inhibitor screening or other biological activity assays may not have been performed for every product. All available data for a given product is shown on the lot-specific Technical Data Sheet.

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.

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.