Anti-Chp1 antibody (ab18191)

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Overview

  • Product name
    Anti-Chp1 antibody
  • Description
    Rabbit polyclonal to Chp1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ChIP, WBmore details
  • Species reactivity
    Reacts with: Schizosaccharomyces pombe
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 900 to the C-terminus of Schizosaccharomyces pombe Chp1.

    Read Abcam's proprietary immunogen policy (Peptide available as ab19117.)

  • Positive control
    • This antibody gave a positive signal in S. pombe whole cell lysate.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab18191 in the following tested applications.

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

Application Abreviews Notes
ChIP Use 2.5µg for 107 cells.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 109 kDa (predicted molecular weight: 109 kDa).

Target

  • Relevance
    Chp1 a chromodomain protein associates with the S. pombe mating type locus and telomeres. Chp1 localization to heterochromatin depends on its chromodomain and the C-terminal domain of the protein. Chp1 and centromeric siRNAs are components of the RITS (RNA-induced initiation of transcriptional gene silencing) complex. The association of RITS with centromeres is linked to Dicer activity, which is required to cleave centromeric transcripts to produce short interfering RNAs that actively recruit components of heterochromatin to centromeres.
  • Cellular localization
    Nuclear
  • Database links
    • Alternative names
      • Chromo domain containing protein 1 antibody
      • Chromodomain protein Chp1 antibody

    Images

    • Western blot - Anti-Chp1 antibody (ab18191)
      Western blot - Anti-Chp1 antibody (ab18191)This image is courtesy of Danesh Moazed's Laboratory, Harvard, Boston.
      All lanes : Anti-Chp1 antibody (ab18191) at 1 µg/ml

      Lane 1 : S. pombe chp1Δ lysate
      Lane 2 : S. pombe wildtype lysate

      Predicted band size: 109 kDa
      Observed band size: 109 kDa
      Additional bands at: 102 kDa (possible degradation product)



      ab18191 specifically recognises Chp1 at 109 kDa in wildtype S. pombe lysate but not in chp1Δ lysate.
    • ChIP - Anti-Chp1 antibody (ab18191)
      ChIP - Anti-Chp1 antibody (ab18191)

      X-ChIP performed with ab18191 on Schizosaccharomyces pombe Cell lysate (whole cell). 5µl antibody were used per 500µl of chromatin/cell lysate according to standard S. pombe ChIP protocols. Protein-A Sepharose was used to recover the ChIPs. In the positive control the researcher specifically induced silencing of the ura4+ gene. For the negative control  the researcher performed multiplex PCR with primers amplifying ura4+ and an un-silenced gene. In addition, act1+ was amplified in a separate PCR reaction.  Chp1 only binds to ura4+ (and only if silencing is induced).

      This image is courtesy of an Abreview submitted by Marc B? on 9 February 2006

      See Abreview

    • Western blot - Anti-Chp1 antibody (ab18191)
      Western blot - Anti-Chp1 antibody (ab18191)
      Anti-Chp1 antibody (ab18191) at 1 µg/ml + S.pombe Whole Cell Lysate at 20 µg

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size: 109 kDa
      Observed band size: 115 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 75&42 kDa. We are unsure as to the identity of these extra bands.

    References

    This product has been referenced in:
    • Jih G  et al. Unique roles for histone H3K9me states in RNAi and heritable silencing of transcription. Nature 547:463-467 (2017). ChIP . Read more (PubMed: 28682306) »
    • Zocco M  et al. The Chp1 chromodomain binds the H3K9me tail and the nucleosome core to assemble heterochromatin. Cell Discov 2:16004 (2016). ChIP . Read more (PubMed: 27462451) »
    See all 13 Publications for this product

    Publishing research using ab18191? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    ChIP abreview for Anti-Chp1 antibody - ChIP Grade

     Excellent
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Schizosaccharomyces pombe Cell lysate - whole cell
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 0 second(s)
    Specification of the cross-linking agent: 3% Formaldehyde
    Detection step
    Semiquantitative PCR
    Positive control
    I specifically induce silencing of the ura4+ gene. I see enrichment for this gene in ChIPs performed with antibodies against H3K9me2, Ago1, Swi6 and, as I show here, Chp1. All these proteins are recruited to transcriptional silent loci as for example centromeric DNA repeats. The Chp1 antibody precipitates chromatin very well (see figure )-it gives me the strongest signal among all the antibodies I used so far.
    Negative control
    I perform multiplex PCR with primers amplifying ura4+ and an un-silenced gene. In addition, act1+ was amplified in a separate PCR reaction. As you can see, Chp1 only binds to ura4+ (and only if silencing is induced).
    Read More

    Dr. Marc B?

    Verified customer

    Submitted Feb 09 2006

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