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Recombinant TET2 (1129-2002) protein

Expressed In

Baculovirus

Protein Species

Human

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Cat No : 31418

Validation Data Gallery

  • Recombinant TET2 (1129-2002) protein gel. Recombinant TET2 was run on a 10% SDS-PAGE gel and stained with Coomassie Blue. MW: 102.7 kDa Purity: >45%

    Recombinant TET2 (1129-2002) protein gel. Recombinant TET2 was run on a 10% SDS-PAGE gel and stained with Coomassie Blue. MW: 102.7 kDa Purity: >45%

  • Recombinant TET2 (1129-2002) activity assay 500 nM of 30 bp dsDNA oligos (sequence: 5’-CAGTAACTGTGGTC/5mC/GGTAACTGACTTGCA-3’) were incubated with different concentrations of TET2 (1129-2002) protein in a 20 μl reaction system containing 50 mM HEPES pH 7.9, 50 μM Fe(NH4)2(SO4)2, 100 μM ascorbate, 100 μM alpha-ketoglutarate, 100 μM ATP, 1 mM TCEP at room temperature for 2 hours. 20 μl reaction product was concentrated to 50 ul, then 1 μl product was spotted onto positively charged nylon membrane. 5hmC antibody (Cat. No. 39769) was used to detect the generation of products.

    Recombinant TET2 (1129-2002) activity assay 500 nM of 30 bp dsDNA oligos (sequence: 5’-CAGTAACTGTGGTC/5mC/GGTAACTGACTTGCA-3’) were incubated with different concentrations of TET2 (1129-2002) protein in a 20 μl reaction system containing 50 mM HEPES pH 7.9, 50 μM Fe(NH4)2(SO4)2, 100 μM ascorbate, 100 μM alpha-ketoglutarate, 100 μM ATP, 1 mM TCEP at room temperature for 2 hours. 20 μl reaction product was concentrated to 50 ul, then 1 μl product was spotted onto positively charged nylon membrane. 5hmC antibody (Cat. No. 39769) was used to detect the generation of products.

Product Information

Expressed In Baculovirus
Protein Species Human

Contents

Recombinant TET2 (1129-2002) protein is supplied in 25 mM HEPESNaOH pH 7.5, 300 mM NaCl, 10% glycerol, 0.04% Triton X-100, and 0.5 mM TCEP.

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. TET2 plays an important role in normal myelopoiesis. Somatic loss-of-function mutations in TET2 gene frequently occur in patients with myeloid malignancies. TET2 loss increased hematopoietic stem cell self-renewal and myeloid transformation. TET2 promotes histone O-GlcNAcylation during gene transcription in embryonic stem cells. In addition, TET2 can be recruited to chromatin by CXXC domain protein IDAX. IDAX downregulates TET2 protein by activating caspase pathway.

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 TET2 (1129-2002) protein that includes amino acids 1129-2002 of human TET2 proteins (accession number NP_001120680.1) was expressed in Sf9 cells and contains an N-terminal 6×His-Tag and FLAG-Tag with a molecular weight of 102.7 kDa.

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.