Anti-Acetylated Proteins antibody - ChIP Grade (ab193)

  ChIP - Acetylated Proteins antibody - ChIP Grade (ab193)

Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 5µl of  ab193 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.
    

  Western blot - Acetylated Proteins antibody - ChIP Grade (ab193)

developed using the ECL technique

Exposure time : 5 minutes

ab193 used in Western Blotting.

Lysate prepared from T47D cells was loaded at 100µg. Cells were pre-treated with either insulin, butyrate, resveratrol or trichostatin A. Primary antibody incubated for 24 hours at a 1/1000 dilution. An HRP-conjugated F(ab)2 secondary antibody was used at a 1/8000 dilution.

Left arrows indicate signal increased by treatment with inhibitors of deacetylases. The nuclear protein increased by butyrate and trichostatin A is probably histone H4 based on its size. One unidentified protein of approximately 65kDa is increased by the treatments in cytoplasm.

Image courtesy of an Abreview submitted by J Stanley.