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.
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.
|Components||24 tests||48 tests||96 tests|
|10X Wash Buffer||1 x 5ml||1 x 10ml||1 x 20ml|
|5-hmC Antibody, 0.6 mg/mL||1 x 25µl||1 x 50µl||1 x 100µl|
|8-Well Assay Strips (with Frame)||3 units||6 units||12 units|
|Adhesive 8-Well Strip Film||3 units||6 units||12 units|
|Antibody Buffer||1 x 4ml||1 x 8ml||1 x 16ml|
|Control DNA, 500 ng/mL||1 x 5µl||1 x 10µl||1 x 20µl|
|Control Primer-Forward, 20 µM||1 x 5µl||1 x 10µl||1 x 20µl|
|Control Primer-Reverse, 20 µM||1 x 5µl||1 x 10µl||1 x 20µl|
|DNA Release Buffer||1 x 7ml||1 x 14ml||1 x 28ml|
|hMeDIP Solution||1 x 3ml||1 x 6ml||1 x 12ml|
|Non-Immune IgG ,0.6 mg/mL||1 x 10µl||1 x 20µl||1 x 40µl|
|Proteinase K, 10 mg/mL||1 x 28µl||1 x 56µl||1 x 112µl|
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.
|Functional Studies||Use at an assay dependent concentration.|