Diagenode

iDeal ChIP-seq kit for Histones

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Catalog Number
Format
Price
C01010059
100 rxns
$1,995.00
Other format

Don’t risk wasting your precious sequencing samples. Diagenode’s validated iDeal ChIP-seq kit has everything you need for a successful ChIP prior to Next-Generation Sequencing.The iDeal ChIP-seq kit is the only kit on the market validated for GAIIx (Illumina®) and PGM™ (Ion Torrent™). Our expertise in ChIP-seq tools allows reproducible and efficient results every time. Now, we also offer the iDeal Library Preparation kit to use in conjunction with the iDeal ChIP-seq kit.

  • Characteristics
    • Validated: Only kit on the market validated for GAIIx (Illumina®) and PGM™ (Ion Torrent™)
    • Proven: Our expertise with ChIP-seq tools enables reproducible and efficient results every time
    • Complete: Kit includes control antibodies, control primer pairs and IPure DNA purification module

    Note: to obtain optimal results, this kit should be used in combination with the DiaMag1.5 - magnetic rack.

    ChIP-seq on cells

    Figure 1A

    Figure 1A. The high consistency of the iDeal ChIP-seq kit on the Ion Torrent™ PGM™ (Life Technologies) and GAIIx (Illumina®)
    ChIP was performed on sheared chromatin from 1 million HelaS3 cells using the iDeal ChIP-seq kit and 1 µg of H3K4me3 positive control antibody. Two different biological samples have been analyzed using two different sequencers - GAIIx (Illumina®) and PGM™ (Ion Torrent™). The expected ChIP-seq profile for H3K4me3 on the GAPDH promoter region has been obtained.
    Image A shows a several hundred bp along chr12 with high similarity of read distribution despite the radically different sequencers. Image B is a close capture focusing on the GAPDH that shows that even the peak structure is similar.

    Perfect match between ChIP-seq data obtained with the iDeal ChIP-seq workflow and reference dataset

    Figure 1B

    Figure 1B. The ChIP-seq dataset from this experiment has been compared with a reference dataset from the Broad Institute. We observed a perfect match between the top 40% of Diagenode peaks and the reference dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset.

    Figure 2

    Figure 2. Efficient and easy chromatin shearing using the Bioruptor® and Shearing buffer iS1 from the iDeal ChIP-seq kit
    Chromatin from 1 million of Hela cells was sheared using the Bioruptor® combined with the Bioruptor® Water cooler (Cat No. BioAcc-cool) during 3 rounds of 10 cycles of 30 seconds “ON” / 30 seconds “OFF” at HIGH power setting (position H). Diagenode 1.5 ml TPX tubes (Cat No. M-50001) were used for chromatin shearing. Samples were gently vortexed before and after performing each sonication round (rounds of 10 cycles), followed by a short centrifugation at 4°C to recover the sample volume at the bottom of the tube. The sheared chromatin was then decross-linked as described in the kit manual and analyzed by agarose gel electrophoresis.

    Figure 3

    Figure 3. Validation of ChIP by qPCR: reliable results using Diagenode’s ChIP-seq grade H3K4me3 antibody, isotype control and sets of validated primers
    Specific enrichment on positive loci (GAPDH, EIF4A2, c-fos promoter regions) comparing to no enrichment on negative loci (TSH2B promoter region and Myoglobin exon 2) was detected by qPCR. Samples were prepared using the Diagenode iDeal ChIP-seq kit. Diagenode ChIP-seq grade antibody against H3K4me3 and the corresponding isotype control IgG were used for immunoprecipitation. qPCR amplification was performed with sets of validated primers.

    ChIP-seq on tissue

    Figure 4A

    Figure 4A. Chromatin Immunoprecipitation has been performed using chromatin from mouse liver tissue, the iDeal ChIP-seq kit for Histones and the Diagenode ChIP-seq-grade H3K4me3 (Cat. No. C15410003) antibody. The IP'd DNA was subsequently analysed on an Illumina® HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in a region surrounding the GAPDH positive control gene.

    Figure 4B

    Figure 4B. The ChIP-seq dataset from this experiment has been compared with a reference dataset from the Broad Institute. We observed a perfect match between the top 40% of Diagenode peaks and the reference dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset.

  • Testimonials

    I have been using Diagenode products to perform ChIP-seq during the last three years and I am very satisfied, with the Bioruptor, the kits and the antibodies. I have used the iDeal ChIP-seq kit for Histones and the iDeal ChIP-seq kit for Transcription Factors with very successful and reproducible results. Once I tried to ChIP histones with a home-made protocol and it worked much worse in comparison with Diagenode kits. In other occasion, I tried a non-Diagenode antibody for a transcription factor and I also got much poor results, however with the Diagenode antibody I always got very nice results. I strongly recommend the use of Diagenode products.

    Dr. Francisca Martinez Real - Development and Disease Research Group - Max Planck Institute for Molecular Genetics, Berlin, Germany
  • Applications
    ChIP-seq
    Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on ... Read more
  • Documents
    Chromatin Immunoprecipitation Brochure BROCHURE
    Whether you are experienced or new to the field of chromatin immunoprecipitation, Diagenode has e...
    Download
    Fast and cost-effective ChIP-sequencing using Diagenode iDeal ChIP-seq kit and antibodies with the Ion Torrent PGM™ sequencer POSTER
    ChIP-seq has become the gold standard for whole-genome mapping of protein-DNA interactions. The g...
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    iDeal ChIP Seq histones kit x24 x100 manual MANUAL
    iDeal ChIP Seq histones kit x24 x100 manual
    Download
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: iDeal ChIP-seq kit for Histones (Diagenode Cat# C01010059). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    The Dynamic Epigenetic Landscape of the Retina During Development, Reprogramming, and Tumorigenesis.
    Aldiri I. et al.
    In the developing retina, multipotent neural progenitors undergo unidirectional differentiation in a precise spatiotemporal order. Here we profile the epigenetic and transcriptional changes that occur during retinogenesis in mice and humans. Although some progenitor genes and cell cycle genes were epigenetically sil...

    Genomic responses of mouse synovial fibroblasts during TNF-driven arthritogenesis greatly mimic those of human rheumatoid arthritis
    Ntougkos E. et al.
    OBJECTIVE: Aberrant activation of synovial fibroblasts (SFs) is a key determinant in the pathogenesis of rheumatoid arthritis (RA). We aimed to produce a map of gene expression and epigenetic changes occurring in this cell type during disease progression in the human TNF-transgenic model of arthritis, and identify ...

    Epigenetically-driven anatomical diversity of synovial fibroblasts guides joint-specific fibroblast functions
    Frank-Bertoncelj M, Trenkmann M, Klein K, Karouzakis E, Rehrauer H, Bratus A, Kolling C, Armaka M, Filer A, Michel BA, Gay RE, Buckley CD, Kollias G, Gay S, Ospelt C
    A number of human diseases, such as arthritis and atherosclerosis, include characteristic pathology in specific anatomical locations. Here we show transcriptomic differences in synovial fibroblasts from different joint locations and that HOX gene signatures reflect the joint-specific origins of mouse and human synov...

    Behavioral, transcriptomic and epigenetic responses to social challenge in honey bees
    Shpigler H.Y. et al.
    Understanding how social experiences are represented in the brain and shape future responses is a major challenge in the study of behavior. We addressed this problem by studying behavioral, transcriptomic and epigenetic responses to intrusion in honey bees. Previous research showed that initial exposure to an intrud...

    DNA methylation heterogeneity defines a disease spectrum in Ewing sarcoma
    Sheffield N.C. et al.
    Developmental tumors in children and young adults carry few genetic alterations, yet they have diverse clinical presentation. Focusing on Ewing sarcoma, we sought to establish the prevalence and characteristics of epigenetic heterogeneity in genetically homogeneous cancers. We performed genome-scale DNA methylation ...

    BMP restricts stemness of intestinal Lgr5(+) stem cells by directly suppressing their signature genes
    Zhen Q. et al.
    The intestinal epithelium possesses a remarkable self-renewal ability, which is mediated by actively proliferating Lgr5+ stem cells. Bone morphogenetic protein (BMP) signalling represents one major counterforce that limits the hyperproliferation of intestinal epithelium, but the exact mechanism remains elusive. Here...

    HMCan-diff: a method to detect changes in histone modifications in cells with different genetic characteristics
    Ashoor H. et al.
    Comparing histone modification profiles between cancer and normal states, or across different tumor samples, can provide insights into understanding cancer initiation, progression and response to therapy. ChIP-seq histone modification data of cancer samples are distorted by copy number variation innate to any cancer...

    Epigenetic Networks Regulate the Transcriptional Program in Memory and Terminally Differentiated CD8+ T Cells
    Rodriguez R.M. et al.
    Epigenetic mechanisms play a critical role during differentiation of T cells by contributing to the formation of stable and heritable transcriptional patterns. To better understand the mechanisms of memory maintenance in CD8 + T cells, we performed genome-wide analysis of DNA methylation, histone marking (acetylated...

    Iterative Fragmentation Improves the Detection of ChIP-seq Peaks for Inactive Histone Marks
    Laczik M. et al.
    As chromatin immunoprecipitation (ChIP) sequencing is becoming the dominant technique for studying chromatin modifications, new protocols surface to improve the method. Bioinformatics is also essential to analyze and understand the results, and precise analysis helps us to identify the effects of protocol optimizati...

    Comprehensive genome and epigenome characterization of CHO cells in response to evolutionary pressures and over time
    Feichtinger J, Hernández I, Fischer C, Hanscho M, Auer N, Hackl M, Jadhav V, Baumann M, Krempl PM, Schmidl C, Farlik M, Schuster M, Merkel A, Sommer A, Heath S, Rico D, Bock C, Thallinger GG, Borth N
    The most striking characteristic of CHO cells is their adaptability, which enables efficient production of proteins as well as growth under a variety of culture conditions, but also results in genomic and phenotypic instability. To investigate the relative contribution of genomic and epigenetic modifications towards...

    Brg1 coordinates multiple processes during retinogenesis and is a tumor suppressor in retinoblastoma
    Aldiri I et al.
    Retinal development requires precise temporal and spatial coordination of cell cycle exit, cell fate specification, cell migration and differentiation. When this process is disrupted, retinoblastoma, a developmental tumor of the retina, can form. Epigenetic modulators are central to precisely coordinating developmen...

    Epigenome mapping reveals distinct modes of gene regulation and widespread enhancer reprogramming by the oncogenic fusion protein EWS-FLI1.
    Tomazou EM, Sheffield NC, Schmidl C, Schuster M, Schönegger A, Datlinger P, Kubicek S, Bock C, Kovar H
    Transcription factor fusion proteins can transform cells by inducing global changes of the transcriptome, often creating a state of oncogene addiction. Here, we investigate the role of epigenetic mechanisms in this process, focusing on Ewing sarcoma cells that are dependent on the EWS-FLI1 fusion protein. We establi...

    TRIM28 Represses Transcription of Endogenous Retroviruses in Neural Progenitor Cells.
    Fasching L, Kapopoulou A, Sachdeva R, Petri R, Jönsson ME, Männe C, Turelli P, Jern P, Cammas F, Trono D, Jakobsson J
    TRIM28 is a corepressor that mediates transcriptional silencing by establishing local heterochromatin. Here, we show that deletion of TRIM28 in neural progenitor cells (NPCs) results in high-level expression of two groups of endogenous retroviruses (ERVs): IAP1 and MMERVK10C. We find that NPCs use TRIM28-mediated hi...

    Centromeric histone H2B monoubiquitination promotes noncoding transcription and chromatin integrity.
    Sadeghi L, Siggens L, Svensson JP, Ekwall K
    Functional centromeres are essential for proper cell division. Centromeres are established largely by epigenetic processes resulting in incorporation of the histone H3 variant CENP-A. Here, we demonstrate the direct involvement of H2B monoubiquitination, mediated by RNF20 in humans or Brl1 in Schizosaccharomyces pom...

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Events

  • Epigenetic Mechanisms in Health and Disease 2017
    Barcelona, Spain
    Oct 25-Oct 26, 2017
  • 35ème Congrès de la Societé Française de Toxicologie Génétique
    Marseille, France
    Oct 26-Oct 27, 2017
  • 7th Australian Epigenetics Conference
    Brisbane, Australia
    Oct 29-Nov 1, 2017
  • Australian Genomic Technologies Association (AGTA 2017 Annual Conference)
    Hobart, Tasmania
    Oct 29-Nov 1, 2017
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