Recombinant KDM1B / LSD2 protein

Aliases: Lysine Demethylase 1B, Flavin-Containing Amine Oxidase Domain-Containing Protein 1
Catalog No: 31479 Format: 20 µg ¥75,000 Buy
Catalog No: 31879 Format: 1 mg ¥648,000 Buy
Expressed In: Baculovirus Protein Species: Human

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Contents

Full length recombinant KDM1B / LSD2 protein containing an N-terminal GST-Tag expressed in Sf9 cells and supplied in 25 mM HEPES, pH 7.5, 120 mM NaCl 20 mM GSH and 5% glycerol. Please refer to product insert upon arrival for lot-specific concentration.

Background

KDM1B / LSD2 (Lysine (K)-Specific Demethylase 1B / Lysine-Specific Histone Demethylase 2) is a histone demethylase that demethylates 'Lys-4' of histone H3, a specific tag for epigenetic transcriptional activation, thereby acting as a corepressor. Required for de novo DNA methylation of a subset of imprinted genes during oogenesis. Demethylates both mono- and di-methylated 'Lys-4' of histone H3. Has no effect on tri-methylated 'Lys-4', mono-, di- or tri-methylated 'Lys-9', mono-, di- or tri-methylated 'Lys-27', mono-, di- or tri-methylated 'Lys-36' of histone H3, or on mono-, di- or tri-methylated 'Lys-20' of histone H4 (by similarity).

Application Notes

Recombinant KDM1B / LSD2 protein is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

Specific Activity: H3K4me2 and H3K4me1 demethylase.

Catalytic Ability: > 600 turnovers / enzyme molecule

Histone Demethylase Assay Conditions: 3.3 μM H3K4me2 (1-27aa) peptide was incubated with different concentrations of KDM1B / LSD2 in reaction buffer containing 50 mM TrisCl, pH 8.0, 0.02% Triton X-100, 50 nM FAD for 1 hours at room temperature. HTRF and MALDI-TOF were used for detection.

Protein Details

Full-length Recombinant KDM1B / LSD2 (accession number XM_005248926.1) was expressed in was expressed in Sf9 cells and contains an N-terminal GST-Tag with an observed molecular weight of 116 kDa. The recombinant protein is >92% pure by SDS-PAGE.

References


This product was used in the following publications:
J. Biol. Chem. (2016). 291(47): 24756-24767. PMID: 27634040.

 

Recombinant KDM1B / LSD2 protein gel.
Recombinant KDM1B / LSD2 run on an 8% SDS-PAGE gel and stained with Coomassie blue.

Recombinant KDM1B / LSD2 protein activity assay.
3.3 uM H3K4me2 (1-21aa) peptide was incubated with 5 nM LSD2 in reaction buffer for 2 hours at room temperature. The reaction product was detected by MALDI-TOF. Single 3.3 uM H3K4me2 peptide was used as negative control.

Catalytic Ability: > 600 turnovers/ enzyme molecule

Recombinant KDM1B / LSD2 protein activity assay.
3.3 μM H3K4me2 (1-27aa) peptide was incubated with KDM1B / LSD2 in reaction buffer for 1 hour at room temperature. KDM1B / LSD2 enzyme was used in a HTRF assay to determine enzyme linearity. Demethylated peptide (H3K4me0) was measured using H3K4me0-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 for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.