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

  • Datasheet
  • SDS
  • Protocol Booklet
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5-hmC standard curve
  • Quantification of 5-hmC content of DNA from various species.

Key features and details

  • Detection method: Colorimetric
  • Platform: Microplate reader
  • Sample type: DNA

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Overview

  • Product name

    Global DNA Hydroxymethylation Assay Kit (5hmc, Colorimetric)
  • Detection method

    Colorimetric
  • Sample type

    DNA
  • Product overview

    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.

  • Notes

    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.

     

     

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 48 tests 1 x 96 tests
    10X Wash Buffer 1 x 14ml 1 x 28ml
    5-hmC Antibody, 1000X 1 x 5µl 1 x 10µl
    8-Well Assay Strips (with Frame) 1 x 6 units 1 x 12 units
    Binding solution 1 x 5ml 1 x 10ml
    Developer Solution 1 x 5ml 1 x 10ml
    Enhancer Solution, 1000X 1 x 5µl 1 x 10µl
    Negative Control containing 0% 5-hmC, 50 µg/ml 1 x 50µl 1 x 100µl
    Positive Control containing 1% 5-hmC, 50 µg/ml 1 x 10µl 1 x 20µl
    Signal Indicator, 1000X 1 x 5µl 1 x 10µl
    Stop Solution 1 x 5ml 1 x 10ml

Associated products

    Images

    • 5-hmC standard curve
      5-hmC standard curve

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

       

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

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (0)

    Publishing research using ab233487? Please let us know so that we can cite the reference in this datasheet.

    ab233487 has not yet been referenced specifically in any publications.

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