Immunoflouroescence analysis of HeLa Cells labelling lysine acetylated proteins with ab22550. Formalin fixed cells were permeabalized with 0.1& Triton X-100 in TBS for 10 mins at room temperature and subsequently blocked with BSA at room temperature for 15 mins. Cells were then probed with ab22550 at 1/100 for 1 hour at room temperature. The secondary used was a DyLight® 488 goat anti-mouse used at 1/400 for 30 minutes at room temperature. Additional counterstains used were F-actin with a DyLight® 554 Phalloidin and Neuclei stained using a Hoechst 33342 conjugate. Image was taken at X20 magnification.

Chromatin Co-Immunoprecipitation (ChIP) analysis  using ab22550 binding acetylated lysines in 10_^E+06 LNCaP cells. Protein binding was detected using real-time PCR.

Positive control: Fold enrichment of ab22550.
Negative Control: Non-specific IgG.

ICC/IF image of ab22550 stained MCF7 cells. The cells were 4% formaldehye fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22550, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.