Recombinant Tet1 (1418-2136) protein

Aliases: LCX, CXXC6, MLL-TET1, TET1-MLL, bA119F7.1
Catalog No: 31417 Format: 20 µg ¥4,040 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

TET (Ten Eleven Translocation) family proteins are cytosine oxygenases that catalyze the conversion of 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC). 5hmC can be further oxidized into 5-formylcytosine (5fC) and 5-carboxylcytosine (5caC) by TET proteins. Methylation at the C5 position of cytosine is an epigenetic modification of the mammalian genome that plays an important role in transcriptional regulation. TET proteins are responsible for initiating the enzymatic deamination process leading to cytosine demethylation.

TET1 preferentially binds to CpG-rich sequences at the promoters of both transcriptionally active and Polycomb-repressed genes and modulates gene silencing induced by cytosine methylation by controlling the levels of 5-mC and 5-hmC at gene promoters. TET1 may have the dual function of repressing the expression of a subset of genes through recruitment of transcriptional repressors to promoters. TET1 is important for stem cell maintenance and specification and is involved in the balance between pluripotency and lineage commitment. TET1 plays an important role in regulating adult hippocampal neurogenesis and cognition

Application Notes

Recombinant TET1 (1418-2136) protein is suitable for use in cytosine oxygenase assays, enzyme kinetics, inhibitor screening, and selectivity profiling.

TET Activity Assay Conditions: 250 nM of 30bp DNA oligos (sequence: 5'-CAGTAACTGTGGTC(mC)GGTAACTGACTTGCA-3', contains a 5mCpG site) was incubated with 100 nM recombinant TET1 (1418-2136) protein in buffer containing 50 mM HEPES, pH 8.0, 100 μM Fe(NH4)2(SO4)2, 2 mM ascorbate, 1 mM alpha-ketoglutarate, 1 mM ATP, 1 mM TCEP at 37C for 1 hr. Activity was detected by Dot-blot.

Protein Details

Recombinant TET1 (1418-2136) protein that includes amino acids 1418-2136 of human TET1 (accession number NP_085128.2) was expressed in a baculovirus expression system and contains an N-terminal 6xHis Tag and FLAG-Tag with a molecular weight of 84.3 kDa. The purity of the protein is >75% by SDS-PAGE.

References

This product was used in the following publications:
Huang, Y. et. al.(2019). Cancer Cell. 35(4):677-691. PMID: 30991027.

 

Recombinant TET1 (1418-2136) protein gel.
Recombinant TET1 (1418-2136) protein was run on an 8% SDS-PAGE gel and stained with Coomassie blue.

Recombinant TET1 (1418-2136) protein activity assay.
250 ng of 30 base oligo (containing 5mCpG) was incubated with 0.3125 µg, 0.625 µg or 1.25 µg TET1 in reaction buffer at 37°C for 1 hour. Sample was concentrated to 5 µl, and 1 µl was spotted onto nylon membrane. Anti-5hmC Ab (Cat#: 39769) was used to detect products.

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 guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.