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 Studies more 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
  • Kits/ Lysates/ Other
  • Kits
  • Epigenetic kits
  • DNA modification and methylation

  • Kits/ Lysates/ Other
  • Kits
  • Epigenetic kits
  • ChIP Kits

  • Epigenetics and Nuclear Signaling
  • DNA methylation
  • Methylated DNA

Applications

EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit images

• 

  Functional Studies - EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit (ab117134)
  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.
• 

  Functional Studies - EpiSeeker hydroxymethylated DNA Immunoprecipitation (hMeDIP) Kit (ab117134)
  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

Protocol Booklet