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Chromatin EasyShear Kit – Ultra Low SDS

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Catalog Number
Format
Order
C01020010
10 chrom. prep./60 rxns
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Download the Manual

Previous name of the kit: Chromatin shearing optimization kit – Ultra Low SDS

A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Ultra Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (<0.1%) is optimized for the preparation of chromatin prior to ChIP on histones. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments suitable for ChIP with the preserved epitopes. The kit is validated for cells and tissues.

Check all Chromatin EasyShear Kits.

Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – Read more

    Characteristics
    • Highly optimized protocol for chromatin preparation prior to ChIP on histones SDS concentration optimized for the workflow for histones
    • Validated for cells and tissue
    • Preserves the epitopes
    • Validated with the Bioruptor ultrasonicator
    • Quality of chromatin sample confirmed by ChIP-seq

    Figure 1. Optimal chromatin shearing profile
    HeLa cells were fixed with formaldehyde for 8 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit – Ultra Low SDS (Cat. No. C01020010). Samples were sonicated for 5-10-15 cycles of 30” ON/30” OFF as indicated with Bioruptor Pico using 1.5 ml Bioruptor microtubes with caps (Cat. No. C30010016) followed by de-crosslinking and DNA purification. The fragment size was assessed using agarose gel electrophoresis. A 100 bp ladder was loaded as the size standard.


    Figure 2. Chromatin precipitation
    Sheared chromatin (obtained with the Chromatin EasyShear Kit – Ultra Low SDS) has been used for immunoprecipitation with H3K4me3 and IgG (negative control) antibodies. Quantitative PCR was performed with positive (GAPDH) and negative (TSH2B) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).

    Cells tested

    The Chromatin EasyShear Kit 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, CD8+ T, Blood vascular endothelial cells, Lymphatic endothelial cells, fibroblasts, K562, MDA-MB231
    • Mouse: C2C12, primary HSPC, synovial fibroblasts, HeLa-S3, FACS sorted cells from embryonic kidneys, macrophages, mesodermal cells, myoblasts, NPC, salivary glands, spermatids, spermatocytes, skeletal muscle stem cells, stem cells, Th2
    • Hamster: CHO
    • Other cell lines/speices: compatible, not tested

    Tissue:

    • Bee – brain
    • Daphnia – whole animal
    • Horse – brain, heart, lamina, liver, lung, skeletal muscles, ovary
    • Human – Erwing sarcoma tumor samples
    • Other tissues: compatible, not tested

    Did you use the Chromatin EasyShear Kit on other cell line / tissue / species? Let us know!

     Documents
    Chromatin EasyShear Kit - Ultra Low SDS MANUAL
    The first critical step of a successful Chromatin immunopre...    		 Download
     Safety sheets
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     Publications

    How to properly cite our product/service in your work

    We strongly recommend using this: Chromatin EasyShear Kit – Ultra Low SDS (Hologic Diagenode Cat# C01020010). Click here to copy to clipboard.

    Using our products or services in your publication? Let us know!

    Disruption of histone acetylation homeostasis reveals multilayered chromatin regulation for transcriptional resiliency
    Venu, Vrinda et al.
    Background: Epigenetic modifications, nucleosome occupancy, and three-dimensional chromatin architecture collectively create a multi-layered, highly interactive regulatory system for controlling genomic functionality. Dysregulation of epigenetic processes leads to a plethora of abnormalities including disease s...

    H3K27me3-mediated epigenetic regulation of TET1 in the eutopic endometrium of women with endometriosis and infertility
    Adamczyk, Magdalena et al.
    Endometriosis is a common gynecological condition associated with chronic inflammation, epigenetic dysregulation, and infertility. The TET1 gene, involved in DNA demethylation, may be regulated by repressive histone modifications such as H3K27me3, but its role in endometriosis remains poorly understood. This study...

    TET2 Deficiency Increases the Competitive Advantage of Hematopoietic Stem Cells through Upregulation of Thrombopoietin Receptor Signaling
    Chan S. et al.
    Ten-Eleven Translocation-2 (TET2) mutations drive the expansion of mutant hematopoietic stem cells (HSCs) in clonal hematopoiesis (CH). However, the precise mechanisms by which TET2 mutations confer a competitive advantage to HSCs remain unclear. Here, through an epigenetic drug screen, we discovered that ...

    Assessment of TET1 gene expression, DNA methylation and H3K27me3level of its promoter region in eutopic endometrium of women withendometriosis and infertility.
    Adamczyk Magdalena et al.
    Endometriosis is the cause of infertility. The eutopic endometrium of women with endometriosis showed an aberrant expression pattern of multitude genes. The role of TET1 protein in the pathogenesis of endometriosis and related infertility is not sufficiently known. Further, knowledge on TET1 transcriptional control ...

    Human papillomavirus type 38 alters wild-type p53 activity to promote cellproliferation via the downregulation of integrin alpha 1 expression.
    Romero-Medina, MC and Venuti, A and Melita, G and Robitaille, A andCeraolo, MG and Pacini, L and Sirand, C and Viarisio, D and Taverniti, Vand Gupta, P and Scalise, M and Indiveri, C and Accardi, R and Tommasino, M
    Tumor suppressors can exert pro-proliferation functions in specific contexts. In the beta human papillomavirus type 38 (HPV38) experimental model, the viral proteins E6 and E7 promote accumulation of a wild-type (WT) p53 form in human keratinocytes (HKs), promoting cellular proliferation. Inactivation of p53 by diff...