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Recombinant Proteins & Enzymes

Recombinant YTHDC1 (325-502) protein

Catalog No: 81100 Format: 100 µg $395 Buy Now
Catalog No: 81800 Format: 1 mg $2,600 Buy Now
Expressed In: E. coli Protein Species: Human

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for Recombinant YTHDC1 (325-502) proteinContents

Recombinant YTHDC1 (325-502) protein is supplied in 25 mM Tris-HCl pH 8.0, 300 mM NaCl, 10% glycerol and 0.5 mM TCEP. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant YTHDC1 (325-502) proteinBackground

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.

for Recombinant YTHDC1 (325-502) proteinApplication Notes

Recombinant YTHDC1 (325-502) 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 YTHDC1 (325-502) protein in 10 µl 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. 

for Recombinant YTHDC1 (325-502) proteinProtein Details

Recombinant YTHDC1 (325-502) protein corresponding to amino acids 325-502 that contains the YTH domain sequence of human YTHDC1 (accession number NP_001317627.1) 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 24.9 kDa.

 
Recombinant YTHDC1 (325-502) SDS PAGE gel

Recombinant YTHDC1 (325-502) SDS PAGE gel
13% SDS-PAGE Coomassie staining
MW: 24.9 kDa
Purity: >90%

HTRF Assay for YTHDC1 (325-502) activity

HTRF for YTHDC1 (325-502) activity
Oligo m6A ssDNA (GTTGG/m6A/CTT) was incubated with different concentrations of YTHDC1 (325-502) 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 Assay for YTHDC1 (325-502) activity

HTRF for YTHDC1 (325-502) activity
Oligo m6A ssDNA (GTTGG/m6A/CTT) was incubated with different concentrations of YTHDC1 (325-502) 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. 

for Recombinant YTHDC1 (325-502) 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 YTHDC1 (325-502) proteinGuarantee

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.

 

Technical Data Sheet

Recombinant YTHDC1 (325-502) protein
 

Data Thumbnails

Recombinant YTHDC1 (325-502) SDS PAGE gel

Recombinant YTHDC1 (325-502) SDS PAGE gel
(Click to enlarge and view details)

HTRF Assay for YTHDC1 (325-502) activity

HTRF for YTHDC1 (325-502) activity
(Click to enlarge and view details)

HTRF Assay for YTHDC1 (325-502) activity

HTRF for YTHDC1 (325-502) activity
(Click to enlarge and view details)