EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit (ab117134)

Overview

  • Product nameEpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) KitSee all hydroxymethylated DNA kits ...
  • Assay time
    3h 00m
  • Species reactivity
    Reacts with: Mouse, Human
  • Product overview

    DNA methylation is a covalent modification of the cytosine ring at the 5' position, 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. The biological important of 5-mC as a major epigenetic modification in phenotype and gene expression has been recognized widely. Quite recently, a novel modified nucleotide, 5-hydroxymethylcytosine (5-hmC), has been detected to be abundant in mouse brain and embryonic stem cells. It is a hydroxylated and methylated form of cytosine. Although it is still uncertain its specific role in epigenetics, it is believed it plays a role in DNA demethylation, chromatin remodeling and gene expression regulation.

    EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit (ab117134) enables the user to enrich hydroxymethated DNA using an antibody specific to hydroxymethylcytosine (5-hmC) to immunoprecipitate hydroxymethylated genomic DNA hMeDIP. The enriched hydroxymethylated fractions can be used in various downstream applications including PCR (hMeDIP-PCR) and microarrays (hMeDIP-chip) for investigating gene-specific hydroxymethylation in cells or tissues.

  • Notes

    Store EMH7 at –20°C away from light.

    Store EMH1, EMH5, EMH6, EMH8, EMH9, EMH10, and 8-Well Assay Strips at 4°C away from light.

    Store remaining components at room temperature away from light.

  • Tested applicationsFunctional Studiesmore details

Properties

  • Storage instructionsPlease refer to protocols.
  • Components 24 tests 48 tests 96 tests
    8-Well Assay Strips (with Frame) 3 units 6 units 12 units
    Adhesive 8-Well Strip Film 3 units 6 units 12 units
    EMH1 (10X Wash Buffer) 1 x 5ml 1 x 10ml 1 x 20ml
    EMH10 (Control Primer-Reverse, 20 µM) 1 x 5µl 1 x 10µl 1 x 20µl
    EMH2 (Antibody Buffer) 1 x 4ml 1 x 8ml 1 x 16ml
    EMH3 (hMeDIP Solution) 1 x 3ml 1 x 6ml 1 x 12ml
    EMH4 (DNA Release Buffer) 1 x 7ml 1 x 14ml 1 x 28ml
    EMH5 (Non-Immune IgG ,0.6 mg/ml) 1 x 10µl 1 x 20µl 1 x 40µl
    EMH6 (5-hmC Antibody, 0.6 mg/ml) 1 x 25µl 1 x 50µl 1 x 100µl
    EMH7 (Control DNA, 500 ng/ml) 1 x 5µl 1 x 10µl 1 x 20µl
    EMH8 (Proteinase K, 10 mg/ml) 1 x 28µl 1 x 56µl 1 x 112µl
    EMH9 (Control Primer-Forward, 20 µM) 1 x 5µl 1 x 10µl 1 x 20µl
  • Research Areas

    Applications

    Our Abpromise guarantee covers the use of ab117134 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    Functional Studies Use at an assay dependent concentration.

    EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit images

    • Sensitive detection of gene-specific hydroxymethylation by hMeDIP-QPCR.

      Human brain DNA (500 ng) was fragmented to 200-600 bps with a sonicator. The fragmented DNA was used for hydroxymethylated DNA enrichment with ab117134. Eluted DNA was analyzed by real time PCR with primers specifically for OCT4 or GAPDH sequences in the promoter regions. Results show that the promoter region is hydroxymethylated in OCT4 but not in GAPDH.
    • Selective enrichment of hydroxymethyated DNA with ab117134.

      50 pg of unmethylated, methylated, and hydroxymethylated DNA control were spiked into fragmented human genomic DNA (500 ng). hMeDIP was processed using 5-hmC antibody and non-immune IgG included in the kit. Eluted DNA was analyzed by real time PCR using control primers included in the kit to detect the presence of spiked control DNA.

    Protocols

    References for EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit (ab117134)

    ab117134 has not yet been referenced specifically in any publications.

    Product Wall

    Both of these kits (ab117133 and ab117135) will immunoprecipitate methylated DNA. The ab117133 Kit uses purified DNA as starting materials while the ab117135 Kit uses cells as starting materials. I can confirm that the immunoprecipitated products are s...

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    Ab117134 is shipped in blue ice at 4°C. Dry ice is not required for this particular kit. However, upon arrival, component EMH7 should be stored at 20°C away from light according to the datasheet instructio...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"