Recombinant YTHDC1 protein

Catalog No: 81176 Format: 20 µg $505 Buy

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Expressed In: Baculovirus Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

YTHDC1 (YTH Domain Containing 1), is a regulator of alternative splicing that specifically recognizes and binds N6-methyladenosine (m6A)-containing RNAs. M6A is a modification present at internal sites of mRNAs and some non-coding RNAs and plays a role in the efficiency of mRNA splicing, processing and stability. YTHDC1 acts as a key regulator of exon-inclusion or exon-skipping during alternative splicing via interaction with mRNA splicing factors SRSF3 and SRSF10. It specifically binds m6A-containing mRNAs and promotes recruitment of SRSF3 to its mRNA-binding elements adjacent to m6A sites, leading to exon-inclusion during alternative splicing. In contrast, interaction with SRSF3 prevents interaction with SRSF10, a splicing factor that promotes exon skipping: this prevents SRSF10 from binding to its mRNA-binding sites close to m6A-containing regions, leading to inhibit exon skipping during alternative splicing. YTHDC1 may also regulate alternative splice site selection.

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 YTHDC1 protein was expressed in a baculovirus expression system as the full length protein (accession number NP_001026902.1) with an N-terminal Flag tag. The molecular weight of the protein is 85.6 kDa

 

Recombinant YTHDC1 protein gel
7.5% SDS-PAGE Coomassie staining
MW: 85.6 kDa
Purity: >85%

Pull-down assay for YTHDC1
120 pmol biotinylated oligos ssDNA (DNA sequence: GTTGG/m6A/CTT) was incubated with 120 pmol YTHDC1 protein in binding system containing 25 mM Tris-HCl pH 8.0, 300 mM NaCl, 5% glycerol, 0.1% Triton-X100 for 4 hours at 4°C. Oligos-YTHDC1 compounds were incubated with streptavidin beads for 1 hour at 4°C, and pull-down products were run on a 7.5% SDS-PAGE gel, stained with Coomassie Blue.

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.