Recombinant JMJD1B / KDM3B protein

Aliases: Lysine Demethylase 3B, Jumonji Domain-Containing Protein 1B, Nuclear Protein 5qNCA
Catalog No: 31429 Format: 20 µg $515 Buy
Catalog No: 31829 Format: 1 mg $4,050 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

Lysine (K)-specific demethylase 3B (KDM3B), also known as Jumonji domain-containing protein 1B (JMJD1B), is a histone demethylase that specifically demethylates lysine 9 of histone H3 (H3K9), thereby playing a central role in defining the histone code. Demethylation of lysine residues generates formaldehyde and succinate. KDM3B may have tumor suppressor activity.

Application Notes

Recombinant JMJD1B / KDM3B is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

Specific Activity: H3K9me2 demethylase.

Catalytic Ability: > 160 turnovers / enzyme molecule

Histone Demethylase Assay Conditions: 3.3 μM H3K9me2 peptide was incubated with different concentrations of recombinant JMJD1B / KDM3B 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 were used for detection.

Protein Details

Recombinant JMJD1B / KDM3B (accession number NP_057688.2) was expressed in Sf9 cells and contains an N-terminal 6xHis Tag and FLAG-Tag with a molecular weight of 195.4 kDa. The recombinant protein is >85% pure by SDS-PAGE.

References

This protein is described in the following references:
Kim JY et al. (2012) Mol Cell Biol 32:2917-33. doi: 10.1128/MCB.00133-12.

 

Recombinant JMJD1B / KDM3B protein gel.
JMJD1B / KDM3B protein was run on an 8% SDS-PAGE gel and stained with Coomassie blue.

Recombinant JMJD1B / KDM3B protein activity assay.
3.3 μM H3K9me2 peptide was incubated with 40 nM JMJD1B / KDM3B in reaction buffer for 2 hours at room temperature. The reaction product was detected by MALDI-TOF. Single 3.3 μM H3K9me2 peptide was used as negative control.

Catalytic Ability: >160 turnovers/ enzyme molecule

Recombinant JMJD1B / KDM3B protein activity assay.
3.3 μM H3K9me2 peptide was incubated with recombinant JMJD1B / KDM3B protein in reaction buffer for 1 hours at room temperature. JMJD1B / KDM3B enzyme was used in a HTRF assay to determine enzyme linearity. Demethylated peptide (H3K9me0) was measured using H3K9me0-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.