Reaction Biology - Model ASH1L (His) - Methyltransferase Assay
From Methyltransferase Proteins
ASH1L is a large , multi-domain protein associated with actively transcribed regions of chromatin, ASH1L is the human homolog of Drosophila Ash1, a Trithorax group protein. Like its counterpart in Drosophila, ASH1L contains a SET histone methyltransferase domain and has been found to play a role in the regulation of Hox gene expression. Although the ASH1L SET domain has been shown in vitro to methylate histone peptides on lysine-4 of histone H3 (H3K4), ASH1L is an H3K36 methyltransferase both in vivo and in vitro with nucleosomes as substrate.
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Recruitment of ASH1L by the ncRNA DBE-T to the chromosome 4q35 locus associated with FSHD (facioscapulohumeral muscular dystrophy) leads to increased H3K36me2 and inappropriate gene derepression at the FSHD locus. In a possible positive feedback loop, ASH1L increases expression of DBE-T itself, suggesting ASH1L’s methyltransferase activity or its interaction with DBE-T as potential therapeutic targets for FSHD. RBC’s ASH1L comprises the catalytic domain (AWS/SET/Post-SET) fused to GST.
ACCESSION #: NM_018489
INCLUDES AMINO ACIDS: 2046-2330
TAG(S): N-terminal His-tag
MW: 35.4 kDa
EXPRESSION SYSTEM: E. coli
SUPPLIED AS: Solution of purified recombinant protein in 100mM HEPES, pH 7.1, 500mM NaCl, 0.5 mM TCEP, 5% (v/v) glycerol
STORAGE: -70°C, Thaw quickly and store on ice before use. The remaining, unused, undiluted enzyme should be snap frozen, for example in a dry/ice ethanol bath or liquid nitrogen. Minimize freeze/thaws if possible, but very low volume aliquots (<5 ul) or storage of diluted enzyme is not recommended.
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