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    products/chip-kits/methylated-dna-immunoprecipitation-medip-kit-dna-ab117133.pdf

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Epigenetics and Nuclear Signaling DNA methylation Methylated DNA
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Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (8)References (11)

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Functional Studies
  • Functional Studies - Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133)
  • Functional Studies - Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133)

Key features and details

  • Assay time: 3 hr

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Overview

  • Product name

    Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA
    See all 5-methylcytosine kits
  • Assay time

    3h 00m
  • Product overview

    Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133) enables the user to enrich methylated DNA 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.


    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.

  • Notes

    NOTE: the size of this kit is based on number of TESTS not SAMPLES. A test simply refers to a single assay well. If you are not sure which size is most suitable for you, contact our Scientific Support Team.

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 24 tests 48 tests
    8-Well Assay Strips (with Frame) 3 units 6 units
    8-Well Strip Caps 3 units 6 units
    Anti-5-Methylcytosine (1 mg/mL) 1 x 25µl 1 x 50µl
    Antibody Buffer 1 x 8ml 1 x 16ml
    Binding Buffer 1 x 5ml 1 x 8ml
    DNA Release Buffer 1 x 2ml 1 x 4ml
    Elution Buffer 1 x 0.6ml 1 x 1.2ml
    F-Collection Tube 30 units 50 units
    F-Spin Column 30 units 50 units
    Normal Mouse IgG (1 mg/mL) 1 x 10µl 1 x 20µl
    Proteinase K (10 mg/mL) 1 x 25µl 1 x 50µl
    Reaction Buffer 1 x 4ml 1 x 8ml
    Wash Buffer 1 x 16ml 2 x 16ml
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA methylation
    • Methylated DNA
    • Kits/ Lysates/ Other
    • Kits
    • Epigenetic kits
    • DNA modification and methylation
    • Kits/ Lysates/ Other
    • Kits
    • Epigenetic kits
    • ChIP Kits
  • Alternative names

    • 5 mC
    • 5 MethylCytosine
    • 5mC

Associated products

  • Related Products

    • Methylated DNA Immunoprecipitation (MeDIP) ChIP Kit (ab117135)
    • Methylated DNA Immunoprecipitation (MeDIP) Kit - Tissue (ab117136)

Images

  • Functional Studies
    Functional StudiesHan B et al., PLoS One, 11(8). Fig 5b. doi: 10.1371/journal.pone.0160358 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    MeDIP was performed using 1μg of DNA with ab117133.  Real-time PCR was carried out with MeDIP DNAs and the frequency of DNA methylation of immunoprecipitated DNA vs input DNA was calculated.

  • Functional Studies - Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133)
    Functional Studies - Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133)
    Captured methylated DNA was used for analyzing methylation level of GAPDH and MLH1 promoter with the use of primers and probes specific to GAPDH and MLH1 promoters, respectively.
  • Functional Studies - Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133)
    Functional Studies - Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133)
    For enrichment of methylated DNA using ab117133, DNA (0.5 ug) isolated from MCF-7 cells was added into the microwell. Methylated DNA was captured by 5-mC antibody prebound to the microwells.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (11)

Publishing research using ab117133? Please let us know so that we can cite the reference in this datasheet.

ab117133 has been referenced in 11 publications.

  • Wang X  et al. N-3 Polyunsaturated Fatty Acid Dehydrogenase Fat-1 Regulates Mitochondrial Energy Metabolism by Altering DNA Methylation in Isolated Cells of Transgenic Cattle. Front Mol Biosci 9:857491 (2022). PubMed: 35517863
  • Gao L  et al. MSTN Mutant Promotes Myogenic Differentiation by Increasing Demethylase TET1 Expression via the SMAD2/SMAD3 Pathway. Int J Biol Sci 16:1324-1334 (2020). PubMed: 32210722
  • Vaher K  et al. Cocaine-induced changes in behaviour and DNA methylation in rats are influenced by inter-individual differences in spontaneous exploratory activity. J Psychopharmacol 34:680-692 (2020). PubMed: 32338111
  • Akbariqomi M  et al. Evaluation and statistical optimization of a method for methylated cell-free fetal DNA extraction from maternal plasma. J Assist Reprod Genet 36:1029-1038 (2019). PubMed: 30820784
  • Wang Z  et al. Insertion of a chimeric retrotransposon sequence in mouse Axin1 locus causes metastable kinky tail phenotype. Mob DNA 10:17 (2019). PubMed: 31073336
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-8 of 8 Abreviews or Q&A

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?

Read More

Abcam community

Verified customer

Asked on Sep 24 2013

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.

Read More

Elisa Thomas

Abcam Scientific Support

Answered on Sep 24 2013

Question

I have just acquired the EpiSeeker methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133) and I am looking for the positive control primers for H19ICR, LAP, XIST or MLH1 and the negative control primers for b-actin or GAPDH.

Could you provide any of those primer sequences?

Read More

Abcam community

Verified customer

Asked on Sep 13 2013

Answer



Here are the sequences for human H19ICR and GAPDH:

H19ICR(234 bp length of amplicons):

F: GAGCCGCACCAGATCTTCAG
R: TTGGTGGAACACACTGTGATCA

GAPDH (110 bp length of amplicons)

F: 5'-ACGTAGCTCAGGCCTCAAGA-3'
R: 5'-GCGGGCTCAATTTATAGAAAC-3'

Read More

Elisa Thomas

Abcam Scientific Support

Answered on Sep 13 2013

Question

Hi, I have another question for you... Our downstream application is the DNA Methylation 3x720K CpG Island Plus RefSeq Promoter Array (Nimblegen). In the user´s guide, Nimblegen recommends a1.5 ug yield (for each sample) for methylated (inmunoprecipitated) DNA, in order to do the hybridization step. How much DNA should I use if I perform the MeDIP with your episeeker kit, in order to get 1.5ug at the end?

Read More

Abcam community

Verified customer

Asked on Oct 02 2012

Answer

Thank you for your inquiry.

With use of this kit, about 40 ng/well of meDNA can be generated from 500 ng of inputgenomic DNA. Thus, at least 20 ug of genomic DNA (in 40 wells) are needed for generating 1.5 ug of methylated DNA.



Based onyour feedback, the WGA method is used for amplification of the yielded methylated DNA from 1-2 well ( about 60-80 ng). The amplified DNA is then used for microarray.



I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

Answered on Oct 02 2012

Question

I'm a little bit confussed about the number of tests and samples for each kit... How many kits do I need to purchase for 60 DNA samples??

Read More

Abcam community

Verified customer

Asked on Sep 28 2012

Answer

Thank you for your inquiry.

You will need 1 well per plate dedicated to the negative control if you're running all at the same time. So for 60 samples you'd probably need to orderone 48 sample kit and one 24 sample kit. The kits are in strip format though, so you can save what you don't use for a later time. This would allow you to do 2 negative controls (1 per kit run) and 70 samples.

I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

Answered on Sep 28 2012

Question

I am just wondering if I could use a previously sonicated,  cleaned and re-precipitated DNA in “ MeDIP” using your EpiSeeker kit. Your protocol calls for the sonication step to be performed in EMD2 reaction buffer. What if I simply add sonicated DNA to EMD2 reaction buffer and heat-denature the DNA at  95oC (Step 4 in the protocol).

Read More

Abcam community

Verified customer

Asked on May 24 2012

Answer

Thank you for contacting us.





The pre-sonicated DNA sample can be used if the DNA is in the buffer(such as TE containing low concentration of salts) that would not interfereantibody binding to the meDNA, or use MC2 to dilute the DNA.



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on May 24 2012

Question

What is the ELution buffer EMD6?

Read More

Abcam community

Verified customer

Asked on May 15 2012

Answer

Thank you for contacting us.
The elution buffer EMD6 used is RNAse/DNAse free water.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.
Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on May 15 2012

Question

I have 24 samples and so ordered the '24 test' kit size for this product. However I'm using the kit now and it is nowapparent that, based on the included protocol,this kit is designed for 12 samples with corresponing input controls. The kits labeling is confusing

Read More

Abcam community

Verified customer

Asked on May 07 2012

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one EpiSeeker methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133).

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Abcam Scientific Support

Answered on May 07 2012

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)

Read More

Abcam community

Verified customer

Asked on Jan 31 2012

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.

Read More

Abcam Scientific Support

Answered on Jan 31 2012

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