Diagenode

ChIL probe (rat, anti-mouse)

目录号
格式
价格
C01060021
12 µl/24 rxns
$600.00

ChIL probe is the key reagent of the ChIL-seq assays, a technique that combines immunostaining, transposase tagging and linear amplification for low-input epigenome/protein binding profiling even from single cells . The ChIL probe (goat, anti-rabbit) comprises a secondary antibody (anti-mouse IgG) conjugated with double-stranded DNA (ChIL DNA) containing a T7 promoter and a primer sequence for the sequencing library preparation, including a mosaic end for Tn5 transposase binding (Fig. 1). Moreover, ChIL probe is labelled with tetramethylrhodamine (TAMRA) for microscopic spatial localization.

The ChIL probe has been validated using Diagenode’s ChIL-seq protocol for cells adapted from the classic ChIL-seq protocol developed by Professors Ohkawa and Kimura in Japan ((Nat Cell Biol 21, 287–296 (2019), Nat Protoc 15, 3334–3360 (2020)). For the in situ transposition reaction Diagenode’s Tagmentase (Tn5 transposase) has been used.

Looking for an anti-rabbit ChIL probe? Go to: ChIL probe (goat, anti-rabbit)

  • ChIL-seq features
    • Sample requirement: 100 – 100K adherent cells
    • Visualization of the spatial localization
    • Chromatin preparation-free method
    • Robust library preparation – by applying an antibody-targeted controlled in situ transposition by Tn5 to link the adaptors
    • Decreased PCR duplicates & increased unique reads due to the use of a linear amplification by the T7 RNA polymerase
    • Capability for massively parallel DNA-sequencing because of the 96-well format
  • Example of ChIL-seq results

    Main statistics

    Table 1. Main Statistics of ChIL-seq experiment.
    This experiment was done on triplicates of 1000 or 100 cells per well (mouse C2C12) for H3K4me3 target and negative control (no primary antibody) according to usual ChIL-seq protocol, using unloaded Tn5 Tagmentase (C01070010).



    Heatmap Figures

    The H3K4me3 modification is associated with active transcription and open chromatin. The heatmaps of the signal 3kb upstream and downstream of each TSS present in the mm10 genome for the data obtained with 1,000 cells and with 100 cells are shown in fig. 1 and fig. 2, respectively. Negative controls are represented as “1stminus” samples (left side) and are compared to positive mH3K4me3 samples (right side).

    Figure 1. Heatmap 3kb upstream and downstream of the TSS - 1,000 cells

    Figure 2. Heatmap 3kb upstream and downstream of the TSS - 100 cells



    IGV figures

    Representative IGV coverage obtained with 1,000 cells and with 100 cells are shown in fig. 3. The first 6 samples relate to 1,000 cells data and the last 6 samples relate to 100 cells data. Negative controls are represented as “1stminus” samples and are compared to positive mH3K4me3 samples.

    Figure 3. Representative IGV figures for H3K4me3 target (GAPDH) - 1,000 and 100 cells.


    Figure 4. Representative immunostaining visualization of H3K4me mark on Hela cells (10K/well, 1µg of antibody, 2h incubation) compared to negative conditions (IgG or no primary antibody condition). DAPI counterstains nuclei, TAMRA (tetramethylrhodamine bound to ChIL probes) stains the target and merge is the combination of both staining.

  •  文档
    Chromatin Brochure BROCHURE
    Whether you are experienced or new to the field of chromatin immunoprecipitation, Diagenode has e...
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    ChIL-seq protocol PROTOCOL
    Download
    ChIL Probe Datasheet (rat, anti-mouse) DATASHEET
    Download
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: ChIL probe (rat, anti-mouse) (Diagenode Cat# C01060021). Click here to copy to clipboard.

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