The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/2000 - 1/10000. Detects a band of approximately 25, 65, 130, 240 kDa (predicted molecular weight: 61 kDa).
Use at 2-5 µg/mg of lysate.
Complex that is 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. The CCR4-NOT complex functions as general transcription regulation complex.
Belongs to the WD repeat HIR1 family. Contains 7 WD repeats.
Active complex is found in G1, S and G2 phases.
Differentially phosphorylated during cell cycle. During mitosis the p60 subunit of inactive CAF-1 is hyperphosphorylated and displaced into the cytosol. Progressivly dephosphorylated from G1 to S and G2 phase. Phosphorylated p60 is recruited to chromatin undergoing DNA repair after UV irradiation in G1, S or G2 phases.
Nucleus. Cytoplasm. DNA replication foci. Cytoplasmic in M phase.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human small cell lung cancer tissue labelling p60 CAF1 with ab7252 at 1/200 (1µg/ml). Detection: DAB.
Immunocytochemistry/ Immunofluorescence - Anti-p60 CAF1 antibody (ab72520)This image is courtesy of an Abreview submitted by Dimitra Kalamida
ab72520 staining p60 CAF1 in human PC3 prostate cancer cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 pH 7.4 for 5 minutes at room temperature and blocked with 5% BSA for 20 minutes at room temperature. Samples were incubated with primary antibody (1/100 in PBS) for 1 hour. A CF568-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.
All lanes : Anti-p60 CAF1 antibody (ab72520) at 0.04 µg/ml
Lane 1 : Whole cell lysate from Hela cells at 50 µg Lane 2 : Whole cell lysate from Hela cells at 15 µg Lane 3 : Whole cell lysate from Hela cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg
Immunoprecipitation/ Western Blot of p60 CAF1.
Lane 1: ab72520 at 3µg/mg whole cell lysate.
Lane 2: Control IgG.
ab72520 at 1µg/ml for WB.
Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded.
Chemiluminescence with an exposure time of 3 seconds.