Recombinant
RabMAb

Recombinant Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)

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Overview

  • Product name

    Anti-p150 CAF1/CAF antibody [EPR5576(2)]
    See all p150 CAF1/CAF primary antibodies

  • Description

    Rabbit monoclonal [EPR5576(2)] to p150 CAF1/CAF

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human p150 CAF1/CAF aa 100-200. The exact sequence is proprietary.

  • Positive control

    • Jurkat, HeLa, HeLa treated with hydroxyurea and K562 whole cell lysates; Human colon and tonsil tissues.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     This product was previously labelled as p150 CAF1

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab126625 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 1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 107 kDa).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt 1/1000 - 1/10000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Core component of the CAF-1 complex, a complex thought to mediate chromatin assembly in DNA replication and DNA repair. Assembles histone octamers onto replicating DNA in vitro. CAF-1 performs the first step of the nucleosome assembly process, bringing newly synthesized histones H3 and H4 to replicating DNA; histones H2A/H2B can bind to this chromatin precursor subsequent to DNA replication to complete the histone octamer. CHAF1A binds to histones H3 and H4. It may play a role in heterochromatin maintenance in proliferating cells by bringing newly synthesized cbx proteins to heterochromatic DNA replication foci (By similarity). The CCR4-NOT complex functions as general transcription regulation complex. Also involved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene.

  • Sequence similarities

    Belongs to the CHAF1A family.

  • Developmental stage

    Active complex is found in G1, S and G2 phases.

  • Domain

    Contains one Pro-Xaa-Val-Xaa-Leu (PxVxL) motif, which is required for interaction with chromoshadow domains. This motif requires additional residues -7, -6, +4 and +5 of the central Val which contact the chromoshadow domain.

  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.

  • Cellular localization

    Nucleus. DNA replication foci.
  • Information by UniProt

  • Database links

    • Alternative names

      • CAF 1 antibody
      • CAF 1 subunit A antibody
      • CAF antibody
      • CAF I 150 kDa subunit antibody
      • CAF I p150 antibody
      • CAF-1 subunit A antibody
      • CAF-I 150 kDa subunit antibody
      • CAF-I p150 antibody
      • CAF1 antibody
      • CAF1A_HUMAN antibody
      • CAF1B antibody
      • CAF1P150 antibody
      • CHAF1A antibody
      • Chromatin assembly factor 1 subunit A antibody
      • Chromatin Assembly Factor 1 Subunit A p150 antibody
      • Chromatin assembly factor I (150 kDa) antibody
      • Chromatin assembly factor I 150 kDa antibody
      • Chromatin assembly factor I p150 subunit antibody
      • DCAF1 antibody
      • hp150 antibody
      • MGC71229 antibody
      • P150 antibody
      see all

    Images

    • Western blot - Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)
      Western blot - Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)
      All lanes : Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate, treated with hydroxyourea
      Lane 3 : HeLa cell line lysate
      Lane 4 : K562 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

      Predicted band size: 107 kDa

    • Flow Cytometry - Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)
      Flow Cytometry - Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)
      Overlay histogram showing HeLa cells stained with ab126625 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab126625, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)

      Formalin-fixed, paraffin-embedded human colon tissue stained for p150 CAF1/CAF with ab126625 (1/100 dilution) in immunohistochemical analysis.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p150 CAF1/CAF antibody [EPR5576(2)] (ab126625)

      Formalin-fixed, paraffin-embedded human tonsil tissue stained for p150 CAF1/CAF with ab126625 (1/100 dilution) in immunohistochemical analysis.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Huang J  et al. Remodeling of Interstrand Crosslink Proximal Replisomes Is Dependent on ATR, FANCM, and FANCD2. Cell Rep 27:1794-1808.e5 (2019). Read more (PubMed: 31067464) »
    • Zheng L  et al. CHAF1A interacts with TCF4 to promote gastric carcinogenesis via upregulation of c-MYC and CCND1 expression. EBioMedicine 38:69-78 (2018). Read more (PubMed: 30449701) »
    See all 7 Publications for this product

    Publishing research using ab126625? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    Western blot abreview for Anti-p150 CAF1 antibody [EPR5576(2)]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (SW480 cells, HCT116 cells)
    Loading amount
    25 µg
    Specification
    SW480 cells, HCT116 cells
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 17 2013

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