Diagenode

True MicroChIP Kit

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Catalog Number
Format
Price
C01010130
(AB-002-0016)
16 rxns
$495.00

The True MicroChIP kit in combination with the MicroPlex Library Preparation™ kit allows for performing ChIP-seq on as few as 10,000 cells. This microChIP-seq assay has been validated with the new generation IP-Star® Compact Automated Workstation. Furthermore Diagenode's high quality ChIP-seq grade antibodies have been used. 

Video article

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

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  • Characteristics
    • 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.

  • Testimonials

    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
  • 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 epigenomic researches, namely to investigate protein-DNA interaction on a ... Read more
    ChIP-qPCR
    Chromatin Immunoprecipitation (ChIP) coupled with quantitative PCR can be used to investigate protein-DNA interaction at known genomic binding sites. if sites are not known, qPCR primers can also be designed against potential regulatory region... Read more
  • Documents
    TrueMicroChIP kit MANUAL
    Diagenode provides the new True MicroChIP kit with optimized reagents and protocol to ena...
    Download
    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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  • Publications

    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!

    MEF2C protects bone marrow B-lymphoid progenitors during stress haematopoiesis
    Wang W et al.
    DNA double strand break (DSB) repair is critical for generation of B-cell receptors, which are pre-requisite for B-cell progenitor survival. However, the transcription factors that promote DSB repair in B cells are not known. Here we show that MEF2C enhances the expression of DNA repair and recombination factors in ...

    Activating Receptor Signals Drive Receptor Diversity in Developing Natural Killer Cells
    Freund J at al.
    It has recently been appreciated that NK cells exhibit many features reminiscent of adaptive immune cells. Considerable heterogeneity exists with respect to the ligand specificity of individual NK cells and as such, a subset of NK cells can respond, expand, and differentiate into memory-like cells in a ligand-specif...

    Loss of Ezh2 synergizes with JAK2-V617F in initiating myeloproliferative neoplasms and promoting myelofibrosis
    Shimizu T et al.
    Myeloproliferative neoplasm (MPN) patients frequently show co-occurrence of JAK2-V617F and mutations in epigenetic regulator genes, including EZH2. In this study, we show that JAK2-V617F and loss of Ezh2 in hematopoietic cells contribute synergistically to the development of MPN. The MPN phenotype induced by JAK2-V6...

    Rebalancing gene haploinsufficiency in vivo by targeting chromatin
    Fulcoli FG et al.
    Congenital heart disease (CHD) affects eight out of 1,000 live births and is a major social and health-care burden. A common genetic cause of CHD is the 22q11.2 deletion, which is the basis of the homonymous deletion syndrome (22q11.2DS), also known as DiGeorge syndrome. Most of its clinical spectrum is caused by ha...

    Persistent Alterations in Microglial Enhancers in a Model of Chronic Pain
    Denk F et al.
    Chronic pain is a common and devastating condition that induces well-characterized changes in neurons and microglia. One major unanswered question is why these changes should persist long after the precipitating injury has healed. Here, we suggest that some of the longer-lasting consequences of nerve injury may be h...

    Osterix and RUNX2 are Transcriptional Regulators of Sclerostin in Human Bone
    Flor M. Pérez-Campo, Ana Santurtún, Carmen García-Ibarbia, María A. Pascual, Carmen Valero, Carlos Garcés, Carolina Sañudo, María T. Zarrabeitia, José A. Riancho
    Sclerostin, encoded by the SOST gene, works as an inhibitor of the Wnt pathway and therefore is an important regulator of bone homeostasis. Due to its potent action as an inhibitor of bone formation, blocking sclerostin activity is the purpose of recently developed anti-osteoporotic treatments. Two bone-specific tra...

    KAT2B Is Required for Pancreatic Beta Cell Adaptation to Metabolic Stress by Controlling the Unfolded Protein Response.
    Rabhi N et al.
    The endoplasmic reticulum (ER) unfolded protein response (UPR(er)) pathway plays an important role in helping pancreatic β cells to adapt their cellular responses to environmental cues and metabolic stress. Although altered UPR(er) gene expression appears in rodent and human type 2 diabetic (T2D) islets, the un...

    Desensitization and incomplete recovery of hepatic target genes after chronic thyroid hormone treatment and withdrawal in male adult mice
    Kenji Ohba, Melvin Khee-Shing Leow, Brijesh Kumar Singh, Rohit Anthony Sinha, Ronny Lesmana,Xiao-Hui Liao, Paul Michael Yen
    Here, we examined changes in hepatic gene expression and serum TH/thyrotropin (TSH) levels in adult male mice treated either with a single T3 (20 g/100 g body weight) injection (acute T3) or daily injections for 14 days (chronic T3) followed by 10 days withdrawal. Chromatin immunoprecipitation analysis of repre...

    Methionine-dependent histone methylation at developmentally important gene loci in mouse preimplantation embryos
    Kudo M, Ikeda S, Sugimoto M, Kume S
    The involvement of specific nutrients in epigenetic gene regulation is a possible mechanism underlying nutrition-directed phenotypic alteration. However, the involvement of nutrients in gene-specific epigenetic regulation remains poorly understood. Methionine has been received attention as a possible nutrient involv...

    Dynamic changes in histone modifications precede de novo DNA methylation in oocytes
    Stewart KR et al.
    Erasure and subsequent reinstatement of DNA methylation in the germline, especially at imprinted CpG islands (CGIs), is crucial to embryogenesis in mammals. The mechanisms underlying DNA methylation establishment remain poorly understood, but a number of post-translational modifications of histones are implicated in...

    The genetic association of RUNX3 with ankylosing spondylitis can be explained by allele-specific effects on IRF4 recruitment that alter gene expression
    Matteo Vecellio, Amity R Roberts, Carla J Cohen, Adrian Cortes, Julian C Knight, Paul Bowness, B Paul Wordsworth
    The authors sought to identify the functional basis for the genetic association of single nucleotide polymorphisms (SNP), upstream of the RUNX3 promoter, with ankylosing spondylitis (AS). They performed conditional analysis of genetic association data and used ENCODE data on chromatin remodelling and transcription f...

    Deep sequencing and de novo assembly of the mouse oocyte transcriptome define the contribution of transcription to the DNA methylation landscape
    Veselovska L, Smallwood SA, Saadeh H, Stewart KR, Krueger F, Maupetit-Méhouas S, Arnaud P, Tomizawa S, Andrews S, Kelsey G
    BACKGROUND: Previously, a role was demonstrated for transcription in the acquisition of DNA methylation at imprinted control regions in oocytes. Definition of the oocyte DNA methylome by whole genome approaches revealed that the majority of methylated CpG islands are intragenic and gene bodies are hypermethylated...

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Events

  • Workshop on Chromatin Proteomics
    Crete, Greece
    Oct 3-Oct 8, 2016
  • 2nd Annual Next Generation Sequencing Congress
    Boston, USA
    Oct 3-Oct 4, 2016
  • XXXIX Reunión anual de la Sociedad de Bioquimica y Biología Molecular de Chile
    Puerto Varas, Chile
    Sep 27-Sep 30, 2016
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