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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.

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Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (AB156912), expandable thumbnail
  • Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (AB156912), expandable thumbnail
  • Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (AB156912), expandable thumbnail

Publications

Key facts

Detection method

Colorimetric

Sample types

Nuclear Extracts, Purified protein

Assay type

Enzyme activity

Reactive species

Plants, Mammals, Fungi

Assay time

5h

Sensitivity

> 20 ng

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

Function

Dioxygenase that plays a key role in active DNA demethylation, by catalyzing the sequential oxidation of the modified genomic base 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC) (PubMed:19372391, PubMed:21496894, PubMed:21778364, PubMed:35798741). In addition to its role in DNA demethylation, plays a more general role in chromatin regulation by recruiting histone modifying protein complexes to alter histone marks and chromatin accessibility, leading to both activation and repression of gene expression (PubMed:33833093). Plays therefore a role in many biological processes, including stem cell maintenance, T- and B-cell development, inflammation regulation, genomic imprinting, neural activity or DNA repair (PubMed:31278917). Involved in the balance between pluripotency and lineage commitment of cells and plays a role in embryonic stem cells maintenance and inner cell mass cell specification. Together with QSER1, plays an essential role in the protection and maintenance of transcriptional and developmental programs to inhibit the binding of DNMT3A/3B and therefore de novo methylation (PubMed:33833093). May play a role in pancreatic beta-cell specification during development. In this context, may function as an upstream epigenetic regulator of PAX4 presumably through direct recruitment by FOXA2 to a PAX4 enhancer to preserve its unmethylated status, thereby potentiating PAX4 expression to adopt beta-cell fate during endocrine lineage commitment (PubMed:35798741). Under DNA hypomethylation conditions, such as in female meiotic germ cells, may induce epigenetic reprogramming of pericentromeric heterochromatin (PCH), the constitutive heterochromatin of pericentromeric regions. PCH forms chromocenters in the interphase nucleus and chromocenters cluster at the prophase of meiosis. In this context, may also be essential for chromocenter clustering in a catalytic activity-independent manner, possibly through the recruitment polycomb repressive complex 1 (PRC1) to the chromocenters (By similarity). During embryonic development, may be required for normal meiotic progression in oocytes and meiotic gene activation (By similarity). Binds preferentially to DNA containing cytidine-phosphate-guanosine (CpG) dinucleotides over CpH (H=A, T, and C), hemimethylated-CpG and hemimethylated-hydroxymethyl-CpG (PubMed:29276034).Isoform 1Dioxygenase that plays a key role in active DNA demethylation (PubMed:28531272). Binds to promoters, particularly to those with high CG content (By similarity). In hippocampal neurons, isoform 1 regulates the expression of a unique subset of genes compared to isoform 2, although some overlap exists between both isoforms, hence differentially regulates excitatory synaptic transmission (By similarity). In hippocampal neuron cell cultures, isoform 1 controls both miniature excitatory postsynaptic current amplitude and frequency (By similarity). Isoform 1 may regulate genes involved in hippocampal-dependent memory, leading to positive regulation of memory, contrary to isoform 2 that may decrease memory (By similarity).Isoform 2Dioxygenase that plays a key role in active DNA demethylation (PubMed:28531272). As isoform 1, binds to promoters, particularly to those with high CG content, however displays reduced global chromatin affinity compared with isoform 1, leading to decreased global DNA demethylation compared with isoform 1 (By similarity). Contrary to isoform 1, isoform 2 localizes during S phase to sites of ongoing DNA replication in heterochromatin, causing a significant de novo 5hmC formation, globally, and more so in heterochromatin, including LINE 1 interspersed DNA repeats leading to their activation (By similarity). In hippocampal neurons, isoform 2 regulates the expression of a unique subset of genes compared to isoform 1, although some overlap between both isoforms, hence differentially regulates excitatory synaptic transmission (By similarity). In hippocampal neuron cell cultures, isoform 2 controls miniature excitatory postsynaptic current frequency, but not amplitude (By similarity). Isoform 2 may regulate genes involved in hippocampal-dependent memory, leading to negative regulation of memory, contrary to isoform 1 that may improve memory (By similarity). In immature and partially differentiated gonadotrope cells, directly represses luteinizing hormone gene LHB expression and does not catalyze 5hmC at the gene promoter (By similarity).

Alternative names

What's included?

96 Test
Components
10X TET Substrate
1 x 20 µL
10X Wash Buffer
1 x 28 mL
8-Well Assay Strips (with Frame)
1 x 12 Unit
Binding Solution
1 x 10 mL
Capture Antibody, 1000 µg/mL
1 x 8 µL
Co-factor 1
1 x 50 µL
Co-factor 2
1 x 50 µL
Co-factor 3
1 x 50 µL
Detection Antibody, 400 µg/mL
1 x 16 µL
Developer Solution
1 x 10 mL
Enhancer Solution
1 x 16 µL
Stop Solution
1 x 6 mL
TET Assay Buffer
1 x 6 mL
TET Assay Standard, 20 µg/mL
1 x 20 µL

Recommended products

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.

Key facts

Detection method

Colorimetric

Sample types

Nuclear Extracts, Purified protein

Assay type

Enzyme activity

Reactive species

Plants, Mammals, Fungi

Assay time

5h

Assay Platform

Microplate reader

Sensitivity

> 20 ng

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

Multi

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

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.

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3 product images

  • Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912), expandable thumbnail

    Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912)

    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.

  • Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912), expandable thumbnail

    Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912)

    Illustrated standard curve generated with the TET assay standard.

  • Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912), expandable thumbnail

    Functional Studies - TET Hydroxylase Activity Quantification Kit (Colorimetric) (ab156912)

    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).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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