Diagenode

iDeal ChIP-seq kit for Transcription Factors

Catalog Number
Format
Price
C01010055
4 chrom. prep./24 IPs
$1,100.00
Other format

DiagenodeのiDeal ChIP-seq for Transcription Factorsキットは、安定した転写因子ChIP-seqの結果を実現するために、特殊化されたソリューションです。 競合する他社のソリューションとは異なり、本キットは高度に最適化されたプロトコルを使用しており、その品質は多数の転写因子実験検証によって裏付けられています。 優れた特異性と感度で高い収量を実現します。

TESTIMONIAL

One of our issues was that we could obtain only a limited number of cells, which is not enough for canonical ChIP-seq protocols. To solve this issue, we used the Diagenode ChIPmentation solution composed of iDeal ChIP-seq Kit for Transcription Factor, TAG Kit for ChIPmentation, and 24 SI for ChIPmentation. We performed ChIPmentation with IP-Star automated system for GATA6 in 2 million GATA6-overxpressing iPS cells. The result showed clear signal/noise ratio and was highly reproducible. This solution also worked in vitro differentiated definitive endoderm cells (data not shown).

Region 1

Region 2

Figure 1. ChIPmentation sequencing profiles for Gata6
Chromatin preparation and immunoprecipitation have been performed on hiPSCs (human induced Pluripotent Stem Cells) overexpressing Gata6 using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Chromatin from 2,000,000 cells was used for the immunoprecipitation in combination with either anti-GATA6 antibody. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).

Takahiro Suzuki, Ph.D., Senior Research Scientist, Cellular Function Conversion Technology Team, RIKEN Center for Integrative Medical Sciences, Yokohama, Japan
  • 特徴
    • Highly optimized protocol for ChIP-seq from cells and tissues
    • Validated for ChIP-seq with multiple transcription factors and non-histone targets
    • Most complete kit available (covers all steps, including the control antibodies and primers)
    • Magnetic beads make ChIP easy, fast and more reproducible
    • Combination with Diagenode ChIP-seq antibodies provides high yields with excellent specificity and sensitivity
    • Purified DNA suitable for any downstream application
    • Easy-to-follow protocol

     

    ChIP-seq on cells

    CTCF Diagenode

    Figure 1. (A) Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors and the Diagenode ChIP-seq-grade CTCF 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.

    CTCF Diagenode

    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.

     

    ChIP-seq figure A

    ChIP-seq figure B

    ChIP-seq figure C

    Figure 2. Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.

     

    ChIP-seq on tissue

    ChIP-seq figure A

    Figure 3A. Chromatin Immunoprecipitation has been performed using chromatin from mouse liver tissue, the iDeal ChIP-seq kit for Transcription Factors and the Diagenode ChIP-seq-grade CTCF 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 Vwf positive control gene.

    Match of the Top40 peaks

    Figure 3B. 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.

  • Species, cell lines, tissues tested

    The iDeal ChIP-seq Kit for Transcription Factors is compatible with a broad variety of cell lines, tissues and species - some examples are shown below. Other species / cell lines / tissues can be used with this kit.

    Cell lines:

    Human: A549, A673, BT-549, CD4 T, HCC1806, HeLa, HepG2, HFF, HK-GFP-MR, ILC, K562, KYSE-180, LapC4, M14, MCF7, MDA-MB-231, MDA-MB-436, RDES, SKNO1, VCaP, U2-OS, ZR-75-1                     

    Mouse: ESC, NPCs, BZ, GT1-7, acinar cells, HSPCs, Th2 cells, keratinocytes

    Cattle: pbMEC, MAC-T

    Other cell lines / species: compatible, not tested

    Tissues:

    Mouse: kidney, heart, brain, iris, liver, limbs from E10.5 embryos

    Horse: liver, brain, heart, lung, skeletal muscle, lamina, ovary

    Other tissues: compatible, not tested

    ChIP on yeast

    The iDeal ChIP-seq kit for TF is compatible with yeast samples. Check out our Application Note presenting an optimized detailed protocol for ChIP on yeast.

    Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? Let us know!

  •  お客様の声

    We were very happy with the method.  It gave good results in the end, and required much smaller samples than we need to reliably perform conventional ChIP-seq.
    In our view, the main advantages of the ChIPmentation kit compared to our conventional ChIP-seq protocol are (most important first):

    • smaller sample requirement,
    • simpler workflow with less that can go wrong,
    • slightly higher resolution and signal: noise ratio.

    ChIPmentation sequencing profiles for p65. Chromatin preparation and immunoprecipitation have been performed on stimulated NIH3T3 cells using the iDeal ChIP-seq kit for TFs (Cat. No. C01010055). Chromatin from 4,000,000 cells was used for the immunoprecipitation in combination with either anti-p65 antibody or IgG. The library preparation was performed with the TAG Kit for ChIPmentation (Cat. No. C01011030) and 24 SI for ChIPmentation (Cat. No. C01011031).

    Researcher from University of Nice-Sophia Antipolis, Nice, France

    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

    I have been doing ChIPs for a very long time and have tried many kits from different sources like Active Motif, Millipore/Upstate, and homemade reagents. The reproducibility and binding efficacy were never optimal for these until a colleague recommended the iDeal ChIP-seq Kit for Transcription Factors from Diagenode. I have done more than one hundred samples of ChIPs and ChIP-seq using this kit. The results are very consistent and the binding efficacy is higher than with all the other methods. I would definitely recommend this ChIP kit from Diagenode to anyone who is trying to do ChIP or ChIP-seq.

    Researcher at Johns Hopkins University, School of Medicine

    There are so many ChIP-related products on the market, but I feel so lucky that I have been using the ones from Diagenode since I started my CHIP-seq project. I have used their iDeal CHIP-seq Kit for Transcription Factors and MicroPlex Library Prep Kit v2. Both of them are fantastic and very reproducible. With the very-well written protocols, you will just be home and dry. Particularly, I want to thank the technical support, who is very patient, knowledgeable and extremely helpful. I would definitely recommend my colleagues to use the CHIP products from Diagenode.

    Dr Kaiyu Lei, Faculty of Medicine, Department of Surgery & Cancer, Imperial College London

    The new Bioruptor® Pico machine has reduced the amount of time spent sonicating Chromatin by a massive amount. Some protocols require quite harsh fixing conditions which meant fragmenting DNA on the old machine was taking many rounds and several times. With the new Bioruptor® Pico machine these sonications were taking just one round of 10 cycles thereby reducing the fragmentation time substantially. Following sonication, I have used the new IDeal ChIP-seq kit. This is a nice straight forward kit that if followed with an appropriate chip validated antibody gave amazing chip-seq results that worked time and again with several different transcription factors. I would recommend both kits for good, consistant chromatin work.

    Dr. Karen Dawson, RNA Biology Group, Cancer Research UK Manchester Institute at the University of Manchester
  •  資料
    Chromatin Brochure BROCHURE
    Whether you are experienced or new to the field of chromatin immunoprecipitation, Diagenode has e...
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    iDeal ChIP-seq Kit for Transcription Factors - Manual MANUAL
    Diagenode’s iDeal ChIP-seq Kit for Transcription Factors is a highly specialized solution f...
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    iDeal ChIP-seq Kit for Transcription Factors -マニュアル マニュアル
    DiagenodeのiDeal ChIP-seq kit for transcription factorsは、強力な転写因子ChIP-seqの結果を得るための高度に特殊化されたソリューションで...
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    Optimize the selection of guide RNA by ChIP to keep CRISPR on-target APPLICATION NOTE
    The mechanisms of target recognition and target specificity of the Cas9 protein is still not comp...
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    Auto ChIPmentation for TFs PROTOCOL
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    Application note - ChIP on yeast APPLICATION NOTE
    The Application Note: "Use of the iDeal® chromatin immunoprecipitation (ChIP) kit for the ide...
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  •  Safety sheets
    iDeal ChIP-seq Kit for Transcription Factors SDS GB en Download
    iDeal ChIP-seq Kit for Transcription Factors SDS US en Download
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