EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit (ab117134)
- Product nameEpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) KitSee all hydroxymethylated DNA kits ...
- Assay time3h 00m
- Species reactivityReacts 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.
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 Studies more details
- 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
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.
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.
References for EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit (ab117134)
ab117134 has not yet been referenced specifically in any publications.