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Recombinant JMJD1B / KDM3B protein
| Aliases: Lysine Demethylase 3B, Jumonji Domain-Containing Protein 1B, Nuclear Protein 5qNCA |
| Catalog No: 31429 | Format: 20 µg | ¥4,280 | Add to Cart |
| Catalog No: 31829 | Format: 1 mg | ¥33,200 | Add to Cart |
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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.
