Recombinant YTHDC2 (1279-1429) protein

Catalog No: 81101 Format: 100 µg $480 Buy
Catalog No: 81801 Format: 1 mg $2,835 Buy

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Expressed In: E. coli 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

Recombinant YTHDC2 (1279-1429) protein is suitable for use in the study of binding assay, inhibitor screening, and selectivity profiling.

Binding Assay Conditions: 3 µM oligo m6A ssDNA (GTTGG/m6A/CTT) was incubated with different concentrations of YTHDC2 (1279-1429) protein in 10 &micrl;l reaction system containing 50 mM HEPES-NaOH pH 7.5, 0.1% BSA for 1 hour, then 10 µl anti-FLAG antibody and SA-XL665 mixture (1:100 dilution in the same 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. 

Protein Details

Recombinant YTHDC2 (1279-1429) protein corresponding to amino acids 1279-1429 that contains the YTH domain sequence of human YTHDC2 (accession number NP_073739.3) was expressed in E. coli cells with an N-terminal 6×His tag and a C-terminal Flag tag. The molecular weight of the protein is 22.2 kDa.

 

Recombinant YTHDC2 (1279-1429) SDS PAGE gel
13% SDS-PAGE Coomassie staining
MW: 22.2 kDa
Purity: >95%

HTRF for YTHDC2 (1279-1429) activity
Oligo m6A ssDNA (GTTGG/m6A/CTT) was incubated with different concentrations of YTHDC2 (1279-1429) protein in a reaction system for 1 hour, then FLAG antibody and SA-XL665 mixture (1:100 dilution in the same 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. 

HTRF for YTHDC2 (1279-1429) activity
Oligo m6A ssDNA (GTTGG/m6A/CTT) was incubated with different concentrations of YTHDC2 (1279-1429) protein in a reaction system for 1 hour, then FLAG antibody and SA-XL665 mixture (1:100 dilution in the same 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. 

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.