The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 107 kDa).
1/10 - 1/100.
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/1000 - 1/10000. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/50 - 1/100.
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.
Belongs to the CHAF1A family.
Active complex is found in G1, S and G2 phases.
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.
Phosphorylated upon DNA damage, probably by ATM or ATR.
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.
Western blot - Anti-p150 CAF1 antibody [EPR5576(2)] (ab126625)
All lanes : Anti-p150 CAF1 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 Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Yang C et al. Regulation of oxidized base damage repair by chromatin assembly factor 1 subunit A. Nucleic Acids Res45:739-748 (2017).
Read more (PubMed: 27794043) »
Xu M et al. Chromatin assembly factor 1, subunit A (P150) facilitates cell proliferation in human hepatocellular carcinoma. Onco Targets Ther9:4023-35 (2016).
Read more (PubMed: 27445493) »