Diagenode

iDeal ChIP-seq kit for Transcription Factors

目录号
格式
价格
C01010054
2 chrom. prep./10 IPs
$600.00
其他格式

Diagenode’s iDeal ChIP-seq Kit for Transcription Factors is a highly validated solution for robust transcription factor and other non-histone proteins ChIP-seq results and contains everything you need for start-to-finish ChIP prior to Next-Generation Sequencing. This complete solution contains all buffers and reagents for cell lysis, chromatin shearing, immunoprecipitation, and DNA purification. In addition, unlike competing solutions, the kit contains positive and negative control antibodies (CTCF and IgG, respectively) as well as positive and negative control PCR primers pairs (H19 and Myoglobin exon 2, respectively) for your convenience and a guarantee of optimal results.

The  iDeal ChIP-seq kit for Transcription Factors is compatible for cells or tissues:

Amount per IP
Cells 4,000,000
Tissues 30 mg

The iDeal ChIP-seq kit is the only kit on the market validated for major sequencing systems. Our expertise in ChIP-seq tools allows reproducible and efficient results every time.

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
  • Characteristics
    • 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, as 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

    Tissues:

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

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

    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!

  •  证明书

    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

    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
  • Additional solutions compatible with iDeal ChIP-seq kit for Transcription Factors

    The Chromatin shearing optimization kit – Low SDS (iDeal Kit for TFs) is the kit compatible with the iDeal ChIP-seq kit for TF, recommended for the optimization of chromatin shearing, a critical step for ChIP.

    ChIP Cross-link Gold should be used in combination with formaldehyde when working with higher order and/or dynamic interactions, for efficient protein-protein fixation.

    For library preparation of immunoprecipitated samples we recommend to use the  MicroPlex Library Preparation Kit - validated for library preparation from picogram inputs.

    ChIP-seq grade antibodies provide high yields with excellent specificity and sensitivity.

    Check the list of available Primer pairs designed for high specificity to specific genomic regions.

    Plus, for our IP-Star Automation users for automated ChIP, check out our automated version of this kit.

  •  文档
    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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