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Global DNA Hydroxymethylation Assay Kit (ab233487) is suitable for detecting global DNA hydroxymethylation levels using DNA isolated from any species including mammals, plants, fungi, bacteria, and viruses in a variety of forms including, but not limited to, cultured cells, fresh and frozen tissues, paraffin-embedded tissues, plasma/serum samples, and body fluid samples.

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Images

Functional Studies - Global DNA Hydroxymethylation Assay Kit (5hmc, Colorimetric) (AB233487), expandable thumbnail
  • Functional Studies - Global DNA Hydroxymethylation Assay Kit (5hmc, Colorimetric) (AB233487), expandable thumbnail

Publications

Key facts

Detection method

Colorimetric

Sample types

DNA

What's included?

1 x 96 Tests
Components
10X Wash Buffer
1 x 28 mL
5-hmC Antibody, 1000X
1 x 10 µL
8-Well Assay Strips (with Frame)
1 x 12 Unit
Binding solution
1 x 10 mL
Developer Solution
1 x 10 mL
Enhancer Solution, 1000X
1 x 10 µL
Negative Control containing 0% 5-hmC, 50 µg/ml
1 x 100 µL
Positive Control containing 1% 5-hmC, 50 µg/ml
1 x 20 µL
Signal Indicator, 1000X
1 x 10 µL
Stop Solution
1 x 10 mL

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Global DNA Hydroxymethylation Assay Kit (ab233487) is suitable for detecting global DNA hydroxymethylation levels using DNA isolated from any species including mammals, plants, fungi, bacteria, and viruses in a variety of forms including, but not limited to, cultured cells, fresh and frozen tissues, paraffin-embedded tissues, plasma/serum samples, and body fluid samples.

Key facts

Detection method

Colorimetric

Sample types

DNA

Assay Platform

Microplate reader

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

The Global DNA Hydroxymethylation Assay Kit (ab233487) is suitable for detecting global DNA hydroxymethylation levels using DNA isolated from any species including mammals, plants, fungi, bacteria, and viruses in a variety of forms including, but not limited to, cultured cells, fresh and frozen tissues, paraffin-embedded tissues, plasma/serum samples, and body fluid samples.

In this assay, DNA is bound to strip-wells that are specifically treated to have a high DNA affinity. The hydroxymethylated fraction of DNA is detected using a 5-hmC mAb-based detection complex in a one-step manner and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer.

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 (5-mC). In somatic cells, 5-mC 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 5-mC is also observed in non-CpG contexts. The biological importance of 5-mC as a major epigenetic modification in phenotype and gene expression has been recognized widely.

5-hydroxymethylcytosine (5-hmC), as a sixth DNA base with functions in transcription regulation, has been detected to be abundant in human and mouse brains and embryonic stem (ES) cells. In mammals, it can be generated by the oxidation of 5-mC, a reaction mediated by the ten-eleven translocation (TET) family of 5-mC-hydroxylases.

5-hmC has been demonstrated to be tissue specific, ranging from undetectable levels in cultured cell lines to 0.6% in human brain tissues, and can be as high as 8% of total DNA in some other species. The biological significance of 5-hmC as an important epigenetic modification in phenotype and gene expression has been recently recognized. For example, global decrease in 5-hmC content (DNA hypohydroxymethylation) has been exhibited in nearly all cancers and has been proposed as a molecular marker and therapeutic target in cancer as well.

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2 product images

  • Functional Studies - Global DNA Hydroxymethylation Assay Kit (5hmc, Colorimetric) (ab233487), expandable thumbnail

    Functional Studies - Global DNA Hydroxymethylation Assay Kit (5hmc, Colorimetric) (ab233487)

    5-hmC standard curve.

    An example of an optimal standard curve generated with 5-hmC standard control.

  • Functional Studies - Global DNA Hydroxymethylation Assay Kit (5hmc, Colorimetric) (ab233487), expandable thumbnail

    Functional Studies - Global DNA Hydroxymethylation Assay Kit (5hmc, Colorimetric) (ab233487)

    Quantification of 5-hmC content of DNA from various species.

    Accurate quantification of 5-hmC content of various DNA samples from different species with the The Global DNA Hydroxymethylation Assay kit (ab233487). The results are closely correlated with those obtained by MS-LC.

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Product protocols

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

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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