Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 5 hr
- Sample type: Cell culture supernatant, Nuclear Extracts, Purified protein, Tissue
- Sensitivity: 20 ng
Product nameTET Hydroxylase Activity Quantification Kit (Colorimetric)
See all TET1 kits
Sample typeCell culture supernatant, Tissue, Nuclear Extracts, Purified protein
Assay typeEnzyme activity
Sensitivity> 20 ng
Assay time5h 00m
Species reactivityReacts with: Plants, Mammals, Fungi
TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912) is suitable for measuring the activity/inhibition of total 5mC hydroxylase TET enzyme using nuclear extracts or purified TET isoforms (TET 1-3) from a broad range of species such as mammalian, plant, fungal, and bacterial, in a variety of forms including, but not limited to cultured cells, fresh and frozen tissues.
DNA methylation occurs by the covalent addition of a methyl group at the 5-carbon of the cytosine ring by DNA methyltransferases, resulting in 5-methylcytosine (5mC). In somatic cells, 5mC is found almost exclusively in the context of paired symmetrical methylation of the dinucleotide CpG, whereas in embryonic stem (ES) cells, a substantial amount of 5mC is also observed in non-CpG contexts. The biological importance of 5mC as amajor epigenetic modification in phenotype and gene expression has been recognized widely.
5-hydroxymethylcytosine (5hmC), as a sixth DNA base with functions in transcription regulation, has been detected to be abundant in human and mouse brain and embryonic stem (ES) cells. In mammals, it can be generated by oxidation of 5mC, a reaction mediated by the ten-eleven translocation (TET) family of 5mC-hydroxylases.
The TET family of 5mC hydroxylases includes TET1, TET2 and TET3. These TET proteins may promote DNA demethylation by binding to CpG-rich regions to prevent unwanted DNA methyltransferase activity, and by converting 5mC to 5hmC and further to 5-carboxylcytosine (5-caC) through hydroxylase activity. It was shown that genomic 5hmC level correlates to TET hydroxylase activity. In addition, TET1 was shown to have dual functions in transcription activation and repression by binding different target genes in ES cells. TET1 is also a fusion partner of the MLL gene in acute myeloid leukemia and is considered an oncoprotein. TET2 is found to be frequently mutated in leukemia and considered to act as tumor suppressor. TET3 has been demonstrated to play a unique role for DNA methylation reprogramming processes in the mammalian zygote. Thus, activating tumor suppressor TET enzymes such as TET2 or inhibiting oncoprotein TET enzymes such as TET1 would be important in benefiting cancer diagnostics and developing new target-based cancer therapeutics.
Storage instructionsPlease refer to protocols.
Components 48 tests 96 tests 10X TET Substrate 1 x 10µl 1 x 20µl 10X Wash Buffer 1 x 14ml 1 x 28ml 8-Well Assay Strips (with Frame) 1 x 6 units 1 x 12 units Binding Solution 1 x 5ml 1 x 10ml Capture Antibody, 1000 µg/mL 1 x 4µl 1 x 8µl Co-factor 1 1 x 25µl 1 x 50µl Co-factor 2 1 x 25µl 1 x 50µl Co-factor 3 1 x 25µl 1 x 50µl Detection Antibody, 400 µg/mL 1 x 8µl 1 x 16µl Developer Solution 1 x 5ml 1 x 10ml Enhancer Solution 1 x 8µl 1 x 16µl Stop Solution 1 x 3ml 1 x 6ml TET Assay Buffer 1 x 3ml 1 x 6ml TET Assay Standard, 20 µg/mL 1 x 10µl 1 x 20µl
FunctionDioxygenase that catalyzes the conversion of methylcytosine (5mC) to 5-hydroxymethylcytosine (hmC). Plays a role in embryonic stem (ES) cell maintenance and inner cell mass (ICM) cell specification, possibly by participating in DNA demethylation. Specifically binds 5mC, a minor base in mammalian DNA found in repetitive DNA elements that is crucial for retrotransposon silencing and mammalian development. 5mC is present in ES cells and is enriched in the brain, especially in Purkinje neurons. The clear function of hmC is still unclear but it could constitute an intermediate component in cytosine demethylation. A role of hmC in DNA demethylation is supported by TET1 function in ES cell maintenance, which is required to prevent NANOG hypermethylation and maintain NANOG expression in ES cells.
Tissue specificityExpressed in fetal heart, lung and brain, and in adult skeletal muscle, thymus and ovary. Not detected in adult heart, lung or brain.
Involvement in diseaseNote=A chromosomal aberration involving TET1 may be a cause of acute leukemias. Translocation t(10;11)(q22;q23) with MLL. This is a rare chromosomal translocation 5' MLL-TET1 3'.
Sequence similaritiesBelongs to the TET family.
Contains 1 CXXC-type zinc finger.
- Information by UniProt
- CXXC 6
- CXXC finger 6
- Nuclear Extraction Kit (ab113474)
- Hydroxymethylated DNA Quantification Kit (Colorimetric) (ab117130)
- Hydroxymethylated DNA Quantification Kit (Fluorometric) (ab117131)
- Hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit (ab117134)
- TET Hydroxylase Activity Quantification Kit (Fluorometric) (ab156913)
Demonstration of high sensitivity and specificity of the TET activity assay achieved by using nuclear extracts with TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912). Nuclear extracts were prepared from Mouse ES cells.
Illustrated standard curve generated with the TET assay standard.
Demonstration of high sensitivity and specificity of the TET1 activity/inhibition assay achieved by using recombinant TET1 with the TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912).
ab156912 has been referenced in 3 publications.
- Shuai L et al. SIRT5 Regulates Brown Adipocyte Differentiation and Browning of Subcutaneous White Adipose Tissue. Diabetes N/A:N/A (2019). PubMed: 31010955
- Awada Z et al. DNA methylome-wide alterations associated with estrogen receptor-dependent effects of bisphenols in breast cancer. Clin Epigenetics 11:138 (2019). PubMed: 31601247
- Lecoutre S et al. Reduced PPAR?2 expression in adipose tissue of male rat offspring from obese dams is associated with epigenetic modifications. FASEB J 32:2768-2778 (2018). PubMed: 29295860