Methylated DNA Immunoprecipitation (MeDIP) ChIP Kit (ab117135)

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Overview

  • Product name

    Methylated DNA Immunoprecipitation (MeDIP) ChIP Kit
    See all 5-methylcytosine kits

  • Sample type

    Adherent cells, Suspension cells

  • Assay time

    4h 00m

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat

  • Product overview

    DNA methylation is a covalent modification of the cytosine ring at the 5' position of a CpG dinucleotide which leads to epigenetic inactivation of genes when found in 5'-CpG-3'dinucleotides within promoters or in the first exon of genes. It has been demonstrated that DNA methylation plays an important role in the regulation of gene expression, tumorigenesis, and other genetic and epigenetic diseases, and that alterations in DNA methylation patterns are associated with cancer.

    Methylated DNA Immunoprecipitation (MeDIP) ChIP Kit (ab117135) enables the user to enrich methylated DNA directly from cells by using an antibody specific to methyl cytosine (5-mC) to immunoprecipitate methylated genomic DNA (MeDIP). The enriched methylated fractions can then be used in various downstream applications including qualitative and quantitative PCR, as well as southern blot and DNA microarray.

Properties

References (1)

Publishing research using ab117135? Please let us know so that we can cite the reference in this datasheet.

ab117135 has been referenced in 1 publication.

  • Alahari S  et al. The von Hippel Lindau tumour suppressor gene is a novel target of E2F4-mediated transcriptional repression in preeclampsia. Biochim Biophys Acta Mol Basis Dis N/A:N/A (2018). PubMed: 30053437

Customer reviews and Q&As

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Question

measuring increased methylation of genes of interest after treatment
samples: from tissue
for ab117135: How should tissue be prepared/used (protocol refers to adherent cells and suspension cells only)

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Answer

I apologize again for the incorrect information given originally.

For tissues we recommend, ab117136.
In comparison, kit ab117135 would be used when starting with cells. When using a DNA sample, please use ab117133 instead.

I hope this information is of general help nevertheless.

Please let me know if you have any further questions.

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Question

I am interesting to analyze the methylation and hydroximethylation status of my samples (adherent cells) and i found our products: ab117133 and ab117135.

Are these kits suitable for RT-PCR analysis?Do the 24 or 48 tests correspond to 24 or 48 single immunoprecipitations?For example, if i have to analyze two different cell type, i have to consume 6 tests (two for 5mc, two for negative control IgG and two for imput DNA)?And finally, I am interesting to hydroximethylation, so do you have similar kit for this modification?If i purchase the anti-5-hydroxymethyl Cytidine antibody (ab106918), i could use this antibody and so different negative control (in place of Anti-5-methylcytosine and mouse IgG) with the same kit?

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Answer

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 suitable to use for RT-PCR.

The 24 or 48 tests correspond to single immunoprecipitations and not to the number of samples that you are able to assay. Therefore, if for one cell type you need to do 6 tests, then you will be able to run 4 cell types with a 24 sample kit. The kits are in strip format though, so you can save what you don't use for a later time.

With regards to a hydroxymethylated DNA kit, we have https://www.abcam.com/episeeker-hydroxymethylated-dna-quantification-kit-colorimetric-ab117130.html, Seeker hydroxymethylated DNA Quantification Kit (Fluorometric) (ab117131) or https://www.abcam.com/episeeker-hydroxymethylated-dna-immunoprecipitation-hmedip-kit-ab117134.html.

The more suitable kit would depend on what type of information you want from your samples. The ab117130 and ab117131 kits will allow you to look at global hydroxymethylation levels of their samples and would tell you the percent of hydroxymethylation of your DNA but would not allow for RT-PCR. The kit ab117134 will allow the you to explore regions of hydroxymethylated DNA by immunoprecipitating/enriching the hydroxymethylated DNA followed by RT-PCR or hMeDIP-chip which is maybe more useful to you.

You can indeed purchase the anti-5-hydroxymethyl cytidine antibody (ab106918) and use it with the 5-mC immunoprecipitation kit since you coat the antibody on the wells.

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