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Recombinant PHF8-l protein

Aliases: JHDM1F; MRXSSD; ZNF422
商品コード: 31462 Format: 20 µg ¥79,000 Buy Now
Expressed In: E. coli Protein Species: Human

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for Recombinant PHF8-l proteinContents

20 µg of recombinant PHF8-l protein expressed in E. coli cells and is supplied in 20 mM Bis-Tris, pH 5.5, 200 mM NaCl, 5% (v/v) glycerol and 1 mM DTT. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant PHF8-l proteinBackground

PHF8 (PHD finger protein 8), also known as Lysine (K)-specific demethylase 7B (KDM7B) and JmjC domain-containing histone demethylation protein 1D-B (JHDM1DB), is a member of the JmjC-containing (Jumonji-C) class of histone demethylase proteins that are involved in the regulation of genome function through the removal of methyl groups from histones. PHF8 has two N-terminal domains, a PHD finger that binds trimethylated lysine 4 of histone H3 (H3K4me3) and a Jumonji domain that demethylates monomethylated H3 Lys9 (H3K9me1), Histone H3 dimethyl Lys9 (H3K9me2), Histone H3 dimethyl Lys27 (H3K27me2) (which are all modifications associated with transcriptional repression) and also Histone H3 dimethyl Lys36 (H3K36me2) via an oxidative pathway that requires the presence of Fe(II) and α-ketoglutarate as cofactors. Horton et al. (2010) demonstrated that, in the presence of H3K4me3, PHF8 shows enhanced demethylation of H3K9me2 on the same peptide. PHF8 enzyme is involved in various cellular processes, including DNA replication and repair and transcriptional regulation. Defects in the Jumonji domain of PHF8 lead to Siderius type X-linked mental retardation (MRXSSD).

PHF8-l is a truncated protein (aa 1-447) that includes the N-terminal domain, including the PHD and the Jumonji catalytic domain of PHF8. The protein contains different segments for both recognizing and removing opposing methyl groups. While the PHD domain binds H3K4me3, an alteration associated with transcriptional activation, the Jumonji (catalytic) domain demethylates H3K9me2, a mechanism associated with transcriptional repression. The enzymes’ catalytic activity and substrate specificity is thereby enhanced through the simultaneous binding of multiple domains. The ability to distinguish the function of individual methyl marks could lead to a marked understanding of the language of the histone code.

for Recombinant PHF8-l proteinApplication Notes

Recombinant PHF8-l is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

for Recombinant PHF8-l proteinProtein Details

Recombinant PHF8-l, that includes amino acids 1-447 of PHF8 (accession number NP_055922) was generated by expressing a GST fusion protein containing residues 1-447 of PHF8 in E. coli cells, followed by affinity purification and cleavage of the GST-Tag with thrombin to produce a protein with an observed molecular weight of 51.533 kDa.

for Recombinant PHF8-l proteinReferences

This protein is described in the following reference:
Horton J et al. (2010) Nature 17:38-43

 
PHF8-l Coomassie gel

PHF8-l protein gel.
PHF8-l (10 μg) run on a 4-20% SDS-PAGE gel and stained with Coomassie blue.

PHF8-l activity assay

PHF8-l activity assay.
Recombinant PHF8-l activity measured using a fluorescence-based demethylation assay.

for Recombinant PHF8-l proteinStorage

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.

for Recombinant PHF8-l proteinGuarantee

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.

 

Technical Data Sheet

Recombinant PHF8-l protein
 

Data Thumbnails

PHF8-l Coomassie gel

PHF8-l Coomassie stained gel.
(Click image to enlarge and see details).

PHF8-l activity assay

PHF8-l activity assay.
(Click image to enlarge and see details).