Recombinant JMJD2B / KDM4B protein

Aliases: Lysine Demethylase 4B, Jumonji Domain-Containing Protein 2B, JHDM3B
Catalog No: 31501 Format: 20 µg ¥88,000 Buy
Catalog No: 31905 Format: 1 mg ¥770,000 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

JMJD2B / KDM4B (JmjC domain-containing histone demethylation protein 3B, Jumonji domain-containing protein 2B, JHDM3B) is a histone demethylase that has been shown to demethylate trimethylated lysine 9 (K9me3) and lysine 36 (K36me3) residues of histone H3, converting these trimethylated histone residues to dimethylated or monomethylated form. KDM4B has no activity for monomethylated H3K9 and H3K36. JMJD2B / KDM4B is physically associated with the MLL2 methyltransferase complex where it serves as a co-factor for ERα-mediated transcriptional activation.

Application Notes

Recombinant JMJD2B / KDM4B protein is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

Histone Demethylase Assay Conditions: 3.3 μM H3K9me3 (aa 1-21) peptide was incubated with different concentrations of recombinant JMJD2B / KDM4B protein in reaction buffer containing 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 for 1 hours at room temperature. HTRF and MALDI-TOF was used for detection.

Protein Details

Recombinant JMJD2B / KDM4B (accession number NP_055830.1) was expressed in Sf9 cells and contains an N-terminal 6xHis Tag and FLAG-Tag with an observed molecular weight of 126.9 kDa. The recombinant protein is >90% pure by SDS-PAGE.

References

This product was used in the following publications:
Cell Death Dis. (2018). 9(10): 1038. PMID: 30305606.

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Recombinant JMJD2B / KDM4B protein gel.
JMJD2B / KDM4B protein was run on an 8% SDS-PAGE gel and stained with Coomassie blue.

Recombinant JMJD2B / KDM4B protein HTRF activity assay.
3.3 μM H3K9me3 peptide was incubated with JMJD2B / KDM4B in reaction buffer for 1 hours at room temperature. JMJD2B / KDM4B enzyme was used in a HTRF assay to determine enzyme linearity. Demethylated peptide (H3K9me2) was measured using H3K9me2-specific antibody.

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.