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Recombinant JARID1A / KDM5A protein

Expressed In

Baculovirus

Protein Species

Human

Cat No : 31431,31831 31431

验证数据展示

  • Recombinant JARID1A / KDM5A protein gel. JARID1A / KDM5A protein was run on a 10% SDS-PAGE gel and stained with Coomassie Blue. MW: 196 kDa Purity: >60%

    Recombinant JARID1A / KDM5A protein gel. JARID1A / KDM5A protein was run on a 10% SDS-PAGE gel and stained with Coomassie Blue. MW: 196 kDa Purity: >60%

  • Recombinant JARID1A / KDM5A protein activity assay. 3 uM H3K4me3 (1-21) peptide was incubated with 100 nM JARID1A / KDM5A in a reaction buffer containing 50 mM HEPES pH 7.5, 0.02% Triton X-100, 100 uM 2-OG, 100 uM ascorbate, 50 uM (NH4)2Fe(SO4)2·6H2O for12 hr. at RT. Then 10 μl anti-H3K4me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

    Recombinant JARID1A / KDM5A protein activity assay. 3 uM H3K4me3 (1-21) peptide was incubated with 100 nM JARID1A / KDM5A in a reaction buffer containing 50 mM HEPES pH 7.5, 0.02% Triton X-100, 100 uM 2-OG, 100 uM ascorbate, 50 uM (NH4)2Fe(SO4)2·6H2O for12 hr. at RT. Then 10 μl anti-H3K4me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

  • Recombinant JARID1A / KDM5A protein activity assay. 3 uM H3K4me3 (1-21) peptide was incubated with 100 nM JARID1A / KDM5A in a reaction buffer containing 50 mM HEPES pH 7.5, 0.02% Triton X-100, 100 uM 2-OG, 100 uM ascorbate, 50 uM (NH4)2Fe(SO4)2·6H2O for12 hr. at RT. Then 10 μl anti-H3K4me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

    Recombinant JARID1A / KDM5A protein activity assay. 3 uM H3K4me3 (1-21) peptide was incubated with 100 nM JARID1A / KDM5A in a reaction buffer containing 50 mM HEPES pH 7.5, 0.02% Triton X-100, 100 uM 2-OG, 100 uM ascorbate, 50 uM (NH4)2Fe(SO4)2·6H2O for12 hr. at RT. Then 10 μl anti-H3K4me2 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at room temperature. HTRF assay was used for detection.

产品信息

Expressed In Baculovirus
Protein Species Human

Contents

Full length recombinant JARID1A / KDM5A protein is supplied in 25 mM HEPES-NaOH pH 7.5, 300 mM NaCl, 10% glycerol, 0.04% Triton X-100, and 0.5 mM TCEP.

背景介绍

Lysine (K)-specific demethylase 5A (KDM5A), also known as Jumonji, AT rich interactive domain 1A (JARID1A), is a histone demethylase that specifically demethylates lysine 4 of histone H3 (H3K4), thereby playing a central role in defining the histone code. KDM5A demethylates trimethylated and dimethylated, but not monomethylated H3K4. It does not demethylate histone H3K9, H3K27, H3K36, H3K79 or H4K20. KDM5A may function to stimulate transcription mediated by nuclear receptors and may be involved in transcriptional regulation of HOX proteins during cell differentiation. Also, KDM5A may be involved in the transcriptional repression of cytokines such as CXCL12. It has been shown to bind directly with the Retinoblastoma (pRb) protein which regulates cell proliferation. KDM5A also interacts with Rhombotin-2 which functions distinctly in erythropoiesis and in T-cell leukemogenesis.

Application Notes

This product was manufactured as described in Protein Details. Where possible, Active Motif has developed functional or activity assays for recombinant proteins. Additional characterization such as enzyme kinetic activity assays, inhibitor screening or other biological activity assays may not have been performed for every product. All available data for a given product is shown on the lot-specific Technical Data Sheet.

Protein Details

Recombinant JARID1A / KDM5A (accession number NP_001036068.1) was expressed in Sf9 and contains an N-terminal FLAG-Tag with a molecular weight of 196.3 kDa. The recombinant protein is >85% pure by SDS-PAGE.

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.