Diagenode

True MicroChIP kit

カタログ番号
フォーマット
価格
C01010130
(AB-002-0016)
16 rxns
$510.00

True MicroChIP kitとMicroPlex Library Preparation™kitを組み合わせることにより、わずか10,000細胞でChIP-seqアッセイが実行できます。このmicroChIP-seqアッセイは、新世代のIP-Star® Compact Automated Workstationで検証されており、さらにはDiagenodeの高品質ChIP-seqグレード抗体が採用されています。

Video article

Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells

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  • 特徴
    • Revolutionary: Only 10,000 cells needed for complete ChIP-seq procedure
    • Reliable and accurate sequencing library preparation from just picogram inputs
    • Validated on studies for histone marks and with the IP-Star® Compact Automated Platform

    ChIP on 10,000 cells

    High efficiency ChIP on 10,000 cells
    ChIP efficiency on 10,000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies H3K4me3 (Cat. No. pAb-003-050), H3K27ac (pAb-174-050), H3K9me3 (pAb-056-050) and H3K27me3 (pAb-069-050). Sheared chromatin from 10,000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    ChIP amplification

    Match between True MicroChIP peaks and the reference dataset

    Reliable detection of enrichments in ChIP-seq
    A: ChIP has been peformed with H3K4me3 antibody, amplification of 17 pg of DNA ChIP'd from 10,000 cells and amplification of 35 pg of DNA ChIP'd from 100,000 cells (control experiment). The IP'd DNA was amplified and transformed into a sequencing-ready preparation for the Illumina plateform with the MicroPlex Library Preparation kit. The library was then analysed on an Illumina® Genome Analyzer. Cluster generation and sequencing were performed according to the manufacturer's instructions.
    B: We observed a perfect match between the top 40% of True MicroChIP 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.

    Cell lysis

    Hela cells were fixed with 1% formaldehyde (for 10 minutes at RT)
    Cell lysis was performed using the Lysis Buffer tL1 of the Diagenode True MicroChIP kit. Samples corresponding to 10,000 cells are sheared during 5 rounds of 5 cycles of 30 seconds “ON” / 30 seconds “OFF” with the Bioruptor® Plus combined with the Bioruptor® Water cooler (Cat No. BioAcc-cool) at HIGH power setting (position H). For optimal results, samples are vortexed before and after performing 5 sonication cycles, followed by a short centrifugation at 4°C. 10 μl of DNA (equivalent to 60,000 cells) are analysed on a 1.5% agarose gel.

  •  お客様の声

    I am working with the True MicroChIP & Microplex Library Preparation Kits and several histone modification antibodies like H3K27ac, H3K4me3, H3K36me3, and H3K27me3. I got always very good and reproducible results for my ChIP-seq experiments.

    Andrea Thiesen, ZMB, Developmental Biology, Prof. Dr. Andrea Vortkamp´s lab, University Duisburg-Essen, Germany
  •  資料
    TrueMicroChIP kit MANUAL
    Diagenode provides the new True MicroChIP kit with optimized reagents and protocol to ena...
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    Chromatin Immunoprecipitation Brochure BROCHURE
    Whether you are experienced or new to the field of chromatin immunoprecipitation, Diagenode has e...
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    True MicroChIP and MicroPlex kits APPLICATION NOTE
    From minuscule amounts to magnificent results: reliable ChIP-seq data from 10,000 cells with the ...
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    ChIP kit results with True MicroChIP kit POSTER
    Chromatin immunoprecipitation coupled with high-throughput sequencing (ChIP-seq) has become the g...
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    Application note: Best Workflow Practices for ChIP-seq Analysis with Small Samples APPLICATION NOTE
    Combined chromatin immunoprecipitation and next-generation sequencing (ChIP-seq) has become the g...
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    Chromatin shearing with the Diagenode One APPLICATION NOTE
     The Diagenode One is a new affordable solution for Chromatin shearing in ChIP analysis for ...
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  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: True MicroChIP kit (Diagenode Cat# C01010130). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Interaction of Sox2 with RNA binding proteins in mouse embryonic stem cells.
    Samudyata , Amaral PP, Engström PG, Robson SC, Nielsen ML, Kouzarides T, Castelo-Branco G
    Sox2 is a master transcriptional regulator of embryonic development. In this study, we determined the protein interactome of Sox2 in the chromatin and nucleoplasm of mouse embryonic stem (mES) cells. Apart from canonical interactions with pluripotency-regulating transcription factors, we identified interactions with...

    Development and epigenetic plasticity of murine Müller glia.
    Dvoriantchikova G, Seemungal RJ, Ivanov D
    The ability to regenerate the entire retina and restore lost sight after injury is found in some species and relies mostly on the epigenetic plasticity of Müller glia. To understand the role of mammalian Müller glia as a source of progenitors for retinal regeneration, we investigated changes in gene expres...

    Whsc1 links pluripotency exit with mesendoderm specification.
    Tian TV, Di Stefano B, Stik G, Vila-Casadesús M, Sardina JL, Vidal E, Dasti A, Segura-Morales C, De Andrés-Aguayo L, Gómez A, Goldmann J, Jaenisch R, Graf T
    How pluripotent stem cells differentiate into the main germ layers is a key question of developmental biology. Here, we show that the chromatin-related factor Whsc1 (also known as Nsd2 and MMSET) has a dual role in pluripotency exit and germ layer specification of embryonic stem cells. On induction of differentiatio...

    The epigenetic basis for the impaired ability of adult murine retinal pigment epithelium cells to regenerate retinal tissue.
    Dvoriantchikova G, Seemungal RJ, Ivanov D
    The epigenetic plasticity of amphibian retinal pigment epithelium (RPE) allows them to regenerate the entire retina, a trait known to be absent in mammals. In this study, we investigated the epigenetic plasticity of adult murine RPE to identify possible mechanisms that prevent mammalian RPE from regenerating retinal...

    Spatial confinement downsizes the inflammatory response of macrophages.
    Jain N, Vogel V
    Macrophages respond to chemical/metabolic and physical stimuli, but their effects cannot be readily decoupled in vivo during pro-inflammatory activation. Here, we show that preventing macrophage spreading by spatial confinement, as imposed by micropatterning, microporous substrates or cell crowding, suppresses late ...

    Automethylation-induced conformational switch in Clr4 (Suv39h) maintains epigenetic stability.
    Iglesias N, Currie MA, Jih G, Paulo JA, Siuti N, Kalocsay M, Gygi SP, Moazed D
    Histone H3 lysine 9 methylation (H3K9me) mediates heterochromatic gene silencing and is important for genome stability and the regulation of gene expression. The establishment and epigenetic maintenance of heterochromatin involve the recruitment of H3K9 methyltransferases to specific sites on DNA, followed by the re...

    Epigenetic inheritance mediated by coupling of RNAi and histone H3K9 methylation.
    Yu R, Wang X, Moazed D
    Histone post-translational modifications (PTMs) are associated with epigenetic states that form the basis for cell-type-specific gene expression. Once established, histone PTMs can be maintained by positive feedback involving enzymes that recognize a pre-existing histone modification and catalyse the same modificati...

    Insulin promoter in human pancreatic β cells contacts diabetes susceptibility loci and regulates genes affecting insulin metabolism.
    Jian X, Felsenfeld G
    Both type 1 and type 2 diabetes involve a complex interplay between genetic, epigenetic, and environmental factors. Our laboratory has been interested in the physical interactions, in nuclei of human pancreatic β cells, between the insulin ( gene and other genes that are involved in insulin metabolism. We have ...

    Retinoid-Sensitive Epigenetic Regulation of the Hoxb Cluster Maintains Normal Hematopoiesis and Inhibits Leukemogenesis.
    Qian P, De Kumar B, He XC, Nolte C, Gogol M, Ahn Y, Chen S, Li Z, Xu H, Perry JM, Hu D, Tao F, Zhao M, Han Y, Hall K, Peak A, Paulson A, Zhao C, Venkatraman A, Box A, Perera A, Haug JS, Parmely T, Li H, Krumlauf R, Li L
    Hox genes modulate the properties of hematopoietic stem cells (HSCs) and reacquired Hox expression in progenitors contributes to leukemogenesis. Here, our transcriptome and DNA methylome analyses revealed that Hoxb cluster and retinoid signaling genes are predominantly enriched in LT-HSCs, and this coordinate regula...

    Contrasting epigenetic states of heterochromatin in the different types of mouse pluripotent stem cells.
    Tosolini M, Brochard V, Adenot P, Chebrout M, Grillo G, Navia V, Beaujean N, Francastel C, Bonnet-Garnier A, Jouneau A
    Mouse embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs) represent naive and primed pluripotency states, respectively, and are maintained in vitro by specific signalling pathways. Furthermore, ESCs cultured in serum-free medium with two kinase inhibitors (2i-ESCs) are thought to be the ground naïve pl...

    Histone Deacetylases 1 and 2 Regulate Microglia Function during Development, Homeostasis, and Neurodegeneration in a Context-Dependent Manner.
    Datta M, Staszewski O, Raschi E, Frosch M, Hagemeyer N, Tay TL, Blank T, Kreutzfeldt M, Merkler D, Ziegler-Waldkirch S, Matthias P, Meyer-Luehmann M, Prinz M
    Microglia as tissue macrophages contribute to the defense and maintenance of central nervous system (CNS) homeostasis. Little is known about the epigenetic signals controlling microglia function in vivo. We employed constitutive and inducible mutagenesis in microglia to delete two class I histone deacetylases, ...

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