The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000. Detects a band of approximately 55 kDa (predicted molecular weight: 55 kDa).
p55/DCAF1 is a core histone-binding subunit that may target chromatin assembly factors, chromatin remodeling factors and histone deacetylases to their histone substrates in a manner that is regulated by nucleosomal DNA. Component of several complexes which regulate chromatin metabolism. These include the chromatin assembly factor 1 (CAF-1) complex, which is required for chromatin assembly following DNA replication and DNA repair.
ChIP - Anti-p55/DCAF1 antibody (ab53616)This image is courtesy of an Abreview submitted by Dr Kat Millen
Chromatin was obtained from Kc167 cells fixed with formaldehyde for 10 min. The ChIP was performed with 30-40 µg of chromatin, ab53616 at 1/300 dilution (incubated with samples for 16h at 4°C) and 20µl of Protein G Agarose beads per ChIP. The immunoprecipitated DNA was quantified by real time PCR (SYBR) . Gene A's enhancer region locus (positive control) shows enrichment when compared with it's coding sequence locus (negative control).
All lanes : Anti-p55/DCAF1 antibody (ab53616) at 1/2000 dilution
Lane 1 : Drosophila embryos 0-3 hours after egg deposition Lane 2 : Drosophila embryos 3-6 hours after egg deposition Lane 3 : Drosophila embryos 6-12 hours after egg deposition Lane 4 : Drosophila embryos 12-18 hours after egg deposition Lane 5 : Drosophila embryos 18-24 hours after egg deposition Lane 6 : Larvae Lane 7 : Pupae Lane 8 : Adults
Lysates/proteins at 50 µg per lane.
Secondary I25 labelled protein A
Predicted band size : 55 kDa Observed band size : 55 kDa
Collins H & Moon NS The components of Drosophila histone chaperone dCAF-1 are required for the cell death phenotype associated with rbf1 mutation. G3 (Bethesda)3:1639-47 (2013).
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Lee H et al. A dual role for the dREAM/MMB complex in the regulation of differentiation-specific E2F/RB target genes. Mol Cell Biol32:2110-20 (2012).
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