ABE290

Anti-TET3

from rabbit, purified by affinity chromatography

• Synonym(s): Methylcytosine dioxygenase TET3
• UNSPSC Code: 12352203
• eCl@ss: 32160702

Properties

• biological source: rabbit
• Quality Level: 100
• antibody form: affinity isolated antibody
• antibody product type: primary antibodies
• clone: polyclonal
• purified by: affinity chromatography
• species reactivity: mouse
• species reactivity (predicted by homology): human (based on 100% sequence homology), rat (based on 100% sequence homology), nonhuman primates (based on 100% sequence homology)
• technique(s): ChIP: suitable (ChIP-seq); immunocytochemistry: suitable; immunoprecipitation (IP): suitable; western blot: suitable
• NCBI accession no.: NP_001274420
• UniProt accession no.: O43151
• shipped in: ambient
• target post-translational modification: unmodified
• Gene Information: human ... TET3(200424)

Description

General description

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.

Specificity

Immunogen sequence is present in TET3 isoforms Tet3FL, Tet3s, and Tet3o spliced isoforms reported by Jin, S.G., et al. (2016). Cell Rep. 14(3):493-505), as well as all additional human and mouse TET3 spliced isoforms reported by UniProt (O43151, Q8BG87). Note that UniProt isoform 1 (O43151-1/human and Q8BG87-1/mouse) corresponds to isoform Tet3s, not Tet3FL (NM_001287491.1/human and JX946278.1/mouse).

Immunogen

KLH-conjugated linear peptide corrsponding to an internal sequence within the N-terminal half of human TET3.

Application

Anti-TET3, Cat. No. ABE290, is a highly specific rabbit polyclonal antibody that targets methylcytosine dioxygenase TET3 and has been tested in Chromatin Immunoprecipitation (ChIP), ChIP-seq, Immunocytochemistry, Immunoprecipitation, and Western Blotting.

Immunoprecipitation Analysis: 10 µg from a representative lot immunoprecipitated the exogenously expressed mouse TET3-GFP fusion protein from 500 µg 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 µg 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 µg 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).

Quality

Evaluated by Western Blotting of mouse TET3 GFP fusion.

Western Blotting Analysis: 2 µg/mL of this antibody detected the exogenously expressed mouse TET3 GFP fusion protein in 10 µg of lysate from transfected mouse embryonic stem cells (mESCs).

Target description

193.7/179.4 kDa (human isoform Tet3FL/Tet3s), 194.8/180.4/185.1 kDa (mouse isoform Tet3FL/Tet3s/Tet3o), 166.7/77.47 kDa (UniProt human isoform 2/3), 76.26 kDa (UniProt mouse isoform 2) calculated. Note that UniProt isoform 1 (O43151-1/human and Q8BG87-1/mouse) corresponds to isoform Tet3s, not Tet3FL (NM_001287491.1/human and JX946278.1/mouse).

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt..

Other Notes

Concentration: Please refer to lot specific datasheet.

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): Not applicable
• Flash Point(C): Not applicable