Recombinant JARID1B / KDM5B (2-751) protein

Aliases: Lysine Demethylase 5B, Cancer/Testis Antigen 31, Retinoblastoma-Binding Protein 2 Homolog 1, Histone Demethylase JARID1B
Catalog No: 31518 Format: 20 µg $410 Buy
Expressed In: Baculovirus Protein Species: Human

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Contents

Recombinant JARID1B / KDM5B (2-751) protein was expressed in Sf9 cells and is supplied in 25 mM HEPES pH 7.5, 300 mM NaCl, 5% glycerol, 0.04% Triton X-100, 0.2 mM TCEP. Please refer to product insert upon arrival for lot-specific concentration.

Background

Lysine (K)-specific demethylase 5B (KDM5B), also known as Jumonji, AT rich interactive domain 1B (JARID1B), is a histone demethylase that specifically demethylates lysine 4 of histone H3 (H3K4), thereby playing a central role in defining the histone code. KDM5B demethylates trimethylated, dimethylated and monomethylated H3K4. It does not demethylate histone H3K9 or H3K27. KDM5B functions as a transcriptional corepressor for FOXG1B and PAX9. KDM5B induces the proliferation of breast cancer cells by repressing tumor suppressor genes such as BRCA1 and HOXA5. In contrast, it may act as a tumor suppressor for melanoma.

Application Notes

Recombinant JARID1B / KDM5B (2-751) is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiing.

HDM Activity Assay Conditions: 3 µM H3K4me3 (1-21aa) peptide was incubated with KDM5B (2-751) protein in reaction buffer including 50 mM HEPES pH 7.5, 0.02% Triton X-100, 100 µM 2-OG, 100 µM ascorbate, 50 µM (NH4)2Fe(SO4)2·6H2O for 1 hour at room temperature. HTRF assay and MALDI-TOF was used for activity detection.

Protein Details

Recombinant JARID1B / KDM5B (2-751) (accession number NP_006609) was expressed in Sf9 cells and contains an N-terminal FLAG-Tag with an observed molecular weight of 100 kDa. The recombinant protein is ≥ 80% pure by SDS- PAGE.

 
Recombinant JARID1B / KDM5B (2-751) SDS-PAGE Gel

Recombinant JARID1B / KDM5B (2-751) was run on a 12.5% SDS-PAGE gel and stained with Coomassie Blue.
MW: 100 kDa
Purity: >80%

HTRF assay for JARID1B / KDM5B (2-751) activity

HTRF assay for JARID1B /KDM5B (2-751) activity

3 µM H3K4me3 (1-21 aa) peptide was incubated with KDM5B (2-751) protein in reaction buffer for 1 hour at room temperature.
Anti-H3K4me2-d2 antibody was used to detect reaction product.

MALDI-TOF for JARID1B / KDM5B (2-751) activity

MALDI-TOF for JARID1B / KDM5B (2-751) activity.

3.3 µM H3K4me3 (1-21 aa) peptide was incubated with 100 nM or 200 nM JARID1B / KDM5B (2-751) protein in reaction buffer for 2 hours at room temperature. MALDI-TOF was used for detection.

Catalytic Ability: > 60 turnovers/ enzyme molecule

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.