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Chromatin EasyShear Kit - Low SDS

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Catalog Number
Format
Order
C01020013
4 chrom. prep./ 24 rxns
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Download the Manual

Previous name of the kit: Chromatin shearing optimization kit – Low SDS (iDeal Kit for TFs)

A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on transcription factors and other non-histone proteins. 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, tissues and FFPE samples.

Check out all of the 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 transcription factors and non-histone proteins
    • SDS concentration optimized for the workflow for TFs
    • Validated for cells, tissue and FFPE samples
    • 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 10 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit - Low SDS (Cat. No. C01020013). 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 – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) 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, 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

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

     Documents
    Chromatin EasyShear Kit - Low SDS MANUAL
    Old name: Chromatin shearing optimization kit - Low SDS (for Transcription Factors) ...    		 Download
     Safety sheets
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     Publications

    How to properly cite our product/service in your work

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

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    Sumathipala D. et al
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