MABE1144/1132/1133 Methylcytosine dioxygenase TET3 (EC 1.14.11.n2; UniProt O43151) is encoded by the TET3 (also known as KIAA0401) gene (Gene ID 200424) in human. TET (ten-eleven translocation) proteins (TET1, TET2, and TET3) oxidize 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC), providing a means for active epigenetic DNA demethylation in mammals. TET proteins also modify genomic thymine residues to 5-hydroxyuracil. TET proteins depend on Fe(II) and 2-oxoglutarate as cofactors for activity, and 2-oxoglutarate is known to inhibit their activity, while ascorbate is shown to stimulate TET-mediated cytosine oxidation. In addition, post-translational modifications (PTMs), such as phosphorylation and O-GlcNAcylation, also play a role in regulating the activity of TET proteins. TET1 and TET2 are found highly expressed in mouse embryonic stem cells (mESCs) and TET3 is known to be upregulated in oocytes and oxidize the silenced paternal pronuclear DNA. High levels of TET proteins and genomic 5-hydroxymethylcytosine in neuronal tissues are also reported. Three TET3 spliced isoforms/variants have been reported, the long or full-length isoform Tet3FL and the short isoform Tet3s are upregulated during embryonic stem cell (ESC) differentiation toward the neuronal lineage, while Tet3o is oocyte-specific and not detected in ESCs or in any other cell types or adult mouse tissues tested, indicating a role of Tet3o in paternal genome oxidation in mouse zygotes. Tet3FL and Tet3o contain the entire Tet3s sequence with additional (but different) N-terminal sequence. Only Tet3FL contains an N-terminal CXXC domain that binds to unmethylated CpGs and exhibits highest affinity toward 5caC. Tet3FL is substantially less active than Tet3o and Tet3s, suggesting that the CXXC domain restricts Tet3FL 5mC oxidation capacity.
Synonyms: Methylcytosine dioxygenase TET3
Application: Immunoprecipitation Analysis: 10 microg from a representative lot immunoprecipitated the exogenously expressed mouse TET3-GFP fusion protein from 500 microg lysate of transfected mouse embryonic stem cells (mESCs).Chromatin Immunoprecipitation-sequencing (ChIP-seq) Analysis: A representative lot detected TET3 chromosome enrichment sites by ChIP-seq analysis (10 microg per ChIP) using chromatin preparation from mouse embryonic stem cells (mESCs) transfected to overexpress mouse TET3 (Courtesy of Reik lab, Epigenetics ISP, Babraham Institute, UK).Immunocytochemistry Analysis: A 1:250 dilution from a representative lot immunostained 4% paraformaldehyde-fixed, 0.5% Triton X-100-permeabilized mouse embryonic stem cells (mESCs) transfected to overexpress mouse TET3 oocyte isoform Tet3o (Courtesy of Reik lab, Epigenetics ISP, Babraham Institute, UK).Immunocytochemistry Analysis: A 1:250 dilution from a representative lot detected the immunoreactivity of TET3 oocyte isoform Tet3o in 4% paraformaldehyde-fixed, 0.5% Triton X-100-permeabilized mouse zygotes (Courtesy of Reik lab, Epigenetics ISP, Babraham Institute, UK).Immunoprecipitation Analysis: 10 microg from a representative lot co-immunoprecipitated endogenous OGT with the exogenously overexpressed mouse TET3 from transfected mouse embryonic stem cells (mESCs) by RIME (Rapid Immunoprecipitation Mass spectrometry of Endogenous proteins) (Courtesy of Reik lab, Epigenetics ISP, Babraham Institute, UK).
Other Notes: Concentration: Please refer to lot specific datasheet.