Overview

  • Product nameEpiSeeker ChIP Kit - One StepSee all ChIP Kit kits ...
  • Assay time
    5h 00m
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, all Mammals
  • Product overview

    Protein-DNA interactions play a critical role for cellular functions such as signal transduction, gene transcription and epigenetic silencing. In mammalian cells, interactions between the DNA-binding proteins and cognate promoter sequences are primary determinants in establishing spatial and temporal expression patterns of gene that affect homeostasis, development, and adaptation.

     

    The EpiSeeker ChIP Kit - One Step (ab117138) enables the user to enrich a chromatin fraction containing specific DNA sequences from isolated chromatin to be able to investigate the interaction between proteins and DNA. For best result, we recommend the use of EpiSeeker Chromatin Extraction Kit (ab117152) for sample preparation.

  • Notes

    Store ECP1, Non-Immune IgG, Anti-RNA Polymerase II, Proteinase K, GAPDH Primer – Forward, GADPH Primer – Reverse, and 8-Well Assay Strips (With 1 Frame) at 4°C away from light.

    Store remaining components at room temperature away from light.

    All components of the kit are stable for 6 months from the date of shipment, when stored properly.

     

    If additional control primers are needed, they are available for purchase as ab171517 (reverse primer) and ab171518 (forward primer).

  • Tested applicationsChIP more details

Properties

  • Storage instructionsStore at +4°C. Please refer to protocols.
  • Components Identifier 48 tests 96 tests
    10X Wash Buffer 1 x 10ml 1 x 20ml
    8-Well Assay Strips (with Frame) 6 units 12 units
    8-Well Strip Caps 6 units 12 units
    96-Well PCR Plate 1 unit 1 unit
    Adhesive Covering Film 1 unit 1 unit
    Anti-RNA Polymerase II (1 mg/ml) 1 x 8µl 1 x 16µl
    ChIP Buffer 1 x 6ml 1 x 12ml
    DNA Release Buffer ECP3 1 x 14ml 1 x 28ml
    GAPDH Primer - Forward (20 µM) 1 x 8µl 1 x 16µl
    GAPDH Primer - Reverse (20 µM) 1 x 8µl 1 x 16µl
    Non-Immune IgG (1 mg/ml) 1 x 10µl 1 x 20µl
    Proteinase K (10 mg/ml) 1 x 55µl 1 x 110µl
    Sonication Frame 1 unit 1 unit
  • Research Areas

    Applications

    Our Abpromise guarantee covers the use of ab117138 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Notes
    ChIP Use at an assay dependent concentration.

    EpiSeeker ChIP Kit - One Step images

    • Chromatin was prepared from HeLa cells following Abcam's X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. ChIP was performed with ab8580 (Histone H3 tri methyl K4) using ab117138. No antibody was added to the blank (yellow).

      The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primes and probes are located in the first Kb of the transcribed region.

    • The analysis of enrichment of RNA polymerase II in GAPDH and MLH1 promoters by the EpiSeeker ChIP Kit - One Step, with chromatin extract prepared from formaldehyde fixed colon cancer cells. Captured DNA was used for analyzing levels of RNA polymerase II enriched in the GAPDH and MLH1 promoters.

    Protocols

    References for EpiSeeker ChIP Kit - One Step (ab117138)

    This product has been referenced in:
    • Onyeagucha BC  et al. S100P/RAGE signaling regulates microRNA-155 expression via AP-1 activation in colon cancer. Exp Cell Res 319:2081-90 (2013). ChIP ; Human . Read more (PubMed: 23693020) »

    See 1 Publication for this product

    Product Wall

    la concentration de la chromatine peut être mesurée par Bradford ou par un assai BCA.

    Notez que nous recommandons d’utiliser entre 5-10 ug de chromatine ce qui correspond à ˜ 0.5 to 1 x 10^6 cellules (la chromatine es...

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    The wells of ab117138 are coated with protein A/G to bind to the antibody(s) that you use. Multiple antibodies can be used in the same well. However, multiple targeted proteins will be captured, which may make the downstream application and analysis di...

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    The size can be checked by directly running the sheared chromatin as seen in the attached file. If DNA will be purified. The following quick method may be useful: a) Use 10 uL sample and add 40 uL H2O b) Reverse cross-link by adding 2 uL of 5 M ...

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    In general, the chromatin is easy to be sonicated to 200-1000 bps. If the chromatin extracts can not be broken down fully by sonication, it may be due to insufficient time length, power intensity, foaming of chromatin solution during sonication etc. th...

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    1. We tested this kit with human samples, but it should also work fine with mouse samples.

    2. the primers included in the kit is only for human GAPDH. The following are the suggested primers specific for mouse GAPDH:


    F: 5'-...

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    Hello and I apologize for the late reply.

    1. The strips are the same for the two
    kits.
    2. The concentration of chromatin can be measured by
    regular protein quantification methods. Histone protein :DNA ratio is
    about 2:1. <...

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    L'anti-RNA pol II fournie dans le kit ab117138 est un monoclonal de souris, clone [CTD4H8] qui reconnait la partie C-terminale de toutes les formes de la polymerase (phosphorylée et non-phosphorylée).

    J'espère qu...

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    Thank you for your reply. I am glad to hear your are able to get the correct band.

    Cross linking should not interfere with the IP; however cross-linking is a time-critical procedure. Cross-linking should generally only be carried out for a f...

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    Thank you for contacting us. I am sorry to hear that you have been experiencing issues when in ChIP using this product. According to the Uniprot information for this protein (http://www.uniprot.org/uniprot/Q80TJ7)this product may recognize the ~1...

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    Thank you for contacting us and your interest in our products.

    The ChIP kit which your customer is interested in used the protein A beads for isolation and would therefore not be suitable for use with a goat polyclonal antibody. We do however...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"