Overview

  • Product name
    Anti-Arp5 antibody - ChIP Grade
  • Description
    Rabbit polyclonal to Arp5 - ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ChIPmore details
  • Species reactivity
    Reacts with: Saccharomyces cerevisiae
  • Immunogen

    Recombinant full length protein Arp 5.

  • Positive control
    • S. cereviseae lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab12099 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 87 kDa (predicted molecular weight: 87 kDa).
ChIP Use at an assay dependent concentration.

Target

  • Relevance
    The budding yeast INO80 complex is a conserved ATP-dependent nucleosome remodeler containing actin-related proteins Arp5 and Arp8. Strains lacking INO80, ARP5, or ARP8 have defects in transcription. Ino80, Arp5, and Arp8 are recruited to an HO-induced DSB, where a phosphorylated form of H2A accumulates. Recruitment of Ino80 is compromised in cells lacking the H2A phosphoacceptor S129. INO80-mediated chromatin remodeling seems to occur directly at DSBs, where it appears to facilitate processing of the lesion.
  • Cellular localization
    Nuclear
  • Database links
    • Alternative names
      • Actin-like protein ARP5 antibody
      • ARP5_YEAST antibody
      • P53946 antibody

    Images

    • Anti-Arp5 antibody - ChIP Grade (ab12099) + S.cerevisiae lysate

      Secondary
      Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

      Predicted band size: 87 kDa
      Observed band size: 90 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 70 kDa (possible cross reactivity)

    • All lanes : Anti-Arp5 antibody - ChIP Grade (ab12099) at 0.36 µg/ml

      Lane 1 : S. cerevisiae whole cell lysate (wild type)
      Lane 2 : S. cerevisiae whole cell lysate (arp5 knockout)
      Lane 3 : S. cerevisiae whole cell lysate (arp8 knockout)
      Lane 4 : S. cerevisiae whole cell lysate (ino80 knockout)
      Lane 5 : S. cerevisiae whole cell lysate (anc1 knockout)
      Lane 6 : S. cerevisiae whole cell lysate (nhp10 knockout)
      Lane 7 : S. cerevisiae whole cell lysate (wild type, treated with MMS)
      Lane 8 : S. cerevisiae whole cell lysate (arp8 knockout, treated with MMS)

      Predicted band size: 87 kDa
      Observed band size: 100 kDa why is the actual band size different from the predicted?



      Using ab12099, bands were present in all S. cerevisiae strains tested except for an arp5 knockout strain. This provides conclusive evidence that the ab12099 recognises Arp5.

    References

    This product has been referenced in:
    • Gowans GJ  et al. INO80 Chromatin Remodeling Coordinates Metabolic Homeostasis with Cell Division. Cell Rep 22:611-623 (2018). WB ; Saccharomyces cerevisiae . Read more (PubMed: 29346761) »
    • Billant O  et al. The dominant-negative interplay between p53, p63 and p73: A family affair. Oncotarget 7:69549-69564 (2016). WB . Read more (PubMed: 27589690) »
    See all 8 Publications for this product

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

    Application
    ChIP
    Detection step
    Other
    Sample
    Saccharomyces cerevisiae Cell lysate - whole cell (Whole cell lysate)
    Specification
    Whole cell lysate
    Negative control
    No Antibody sample
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 20 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 37% Formaldehyde
    Positive control
    Sample after 1 hour of DNA double strand break

    Mr. Arvind Panday

    Verified customer

    Submitted Jul 09 2014

    Application
    Western blot
    Loading amount
    3.75e+006 cells
    Gel Running Conditions
    Reduced Denaturing (NUPAGE precasted Invitrogen 10% SDS-PAGE)
    Sample
    Schizosaccharomyces pombe Cell lysate - whole cell (whole cell extract)
    Specification
    whole cell extract
    Blocking step
    0.5% BSA + 5% non fat milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Jul 17 2013

    Answer

    Thank you for contacting us. I have received the following information from the lab: Ab12099 (ARP5): Unfortunately, the full WB image showing the knock-out experiment is not on file, thus we are not able to determine if there were additional bands on the blot. We believe the testing may have been done by a collaborator a while ago, but currently do not have information to confirm that. We have not tested for cross reactivity with Arp4 or Arp8. The immunogen is the full length recombinant protein, thus we do not know where the epitope is located. The sequence homologies for the Arp5 yeast protein are as follows: Arp5-Arp4 = 21-30% (with lots of gaps) Arp5-Arp8 = very poor (30-53% for very short stretches, have lots of gaps) Thus, crossreactivity is quite unlikely. Usually a homology of 85% or higher is likely for cross reactivity, but not as low as 30%. Ab12098 (ARP8): We have no information on whether this antibody crossreacts with Arp4 and Arp5. The immunogen for this antibody is the full length recombinant protein. Again, we do not know the epitope region as it has not been mapped. The sequence homologies for the Arp5 yeast protein are as follows: Arp8-Arp4 = very poor (22-38% for short stretches, have lots of gaps) Arp8-Arp5 = very poor (30-53% for short stretches, have lots of gaps) Again, crossreactivity is quite unlikely as stated above. The expected MW of the unprocessed precursors for each yeast protein are Arp5 = 87 kDa Arp8 = 100 kDa Arp4 = 55 kDa Any other bands of different MW could be non-specific bands (but not Arp4,5 or 8). Unfortunately, we do not send out small aliquots as this is operationally not feasible for us. However, we do guarantee all our products for tested species and applications as listed on the datasheet. The guarantee is valid for 6 months after a purchase. For more details about our Abpromise guarantee please check this link: https://www.abcam.com/abpromise. Should the antibodies not give you satisfactory results, we will be able to replace, credit or refund them. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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